. Author manuscript; available in PMC: 2022 Aug 1.

Published in final edited form as: Hum Mutat. 2021 Jun 24;42(8):903–946. doi: 10.1002/humu.24238

Table 2.

Summary of data for mOLF-resident myocilin (Refseq NM_000261.2) variants predicted to be benign, pathogenic, or likely pathogenic.

A. Benign
Variant	Clinical Data	Cell Assays^a	Thermal Stability	Structure Analysis	Additional Variants	Other

p.Thr293Lys c.878C>A	Diagnosis ● POAG (Alward et al., 1998; Alward et al., 2002; Ennis et al., 2010; Faucher et al., 2002; Fingert et al., 1999; Millá et al., 2013; Williams-Lyn et al., 2000) ● Exfoliation Syndrome (Alward et al., 2002) ● NTG (Weisschuh, Neumann, Wolf, Wissinger, & Gramer, 2005) ● OHT (Faucher et al., 2002) ● PACG (X. Huang et al., 2014) ● Control Group (Faucher et al., 2002) Country/Ethnicity ● German ● Spanish ● Caucasian ● African American ● Chinese ● English ● Canadian Family History of POAG/JOAG? ● Yes (Millá et al., 2013) ● No (Ennis et al., 2010; Faucher et al., 2002; X. Huang et al., 2014; Williams-Lyn et al., 2000) gnomAD? ● Allele count 157, allele frequency 5.6e-4 ● European, Ashkenazi, Latino	Secretion Assay ● 37 °C ○ +++ (Gobeil, Letartre, & Raymond, 2006) ● 30 °C ○ n/a Triton X-100 Solubility Assay ● n/a	● WT-like ○ T_m=52.9 °C (Donegan et al., 2015)	Location ● Surface exposed ● Internal loop of Blade C Interactions ● None Predicted consequence of substitution ● Tolerated	● p.Thr293Met (c.878C>T) ○ gnomAD: Allele count 3, allele frequency 1.21e-5 ○ Tolerated ● p.Thr293Ala (c.877A>G) ○ gnomAD: Allele count 1, allele frequency 4.02e-6 ○ Tolerated	n/a
p.Val329Met c.985G>A	Diagnosis ● JOAG (Svidnicki et al., 2018) ● POAG (Fingert et al., 1999; Liu et al., 2012; Shimizu et al., 2000) ● Control Group (Fingert et al., 1999) Country/Ethnicity ● African American ● Brazilian Family History of POAG/JOAG? ● Yes (Shimizu et al., 2000) ● gnomAD? ● Allele count 72, allele frequency 2.55e-4 ○ African/African American	Secretion Assay ● 37 °C ○ n/a ● 30 °C ○ n/a Triton X-100 Solubility Assay ● Soluble(Shimizu et al., 2000)	● Slightly destabilized, mouse WT-like (Patterson-Orazem et al., 2019) ○ T_m=49.6 °C (Donegan et al., 2015)	Location ● Buried ● Innermost strand of Blade B Interactions ● Hydrophobic pocket with Blade A Predicted consequence of substitution ● Tolerated, rearrangement likely ● Met is residue in D. rerio myocilin	n/a	n/a
p.Glu352Lys c.1054G>A	Diagnosis ● POAG (Alward et al., 1998; Faucher et al., 2002; Fingert et al., 1999; Liu et al., 2012; Whigham et al., 2011; Williams-Lyn et al., 2000) ● Control Group (Alward et al., 1998; Fingert et al., 1999; Liu et al., 2012; Svidnicki et al., 2018; Whigham et al., 2011) ● PCG (Millá et al., 2013) Country/Ethnicity ● Caucasian ● African American ● African Caribbean ● Canadian ● Black South African ● Spanish ● Brazilian Family History of POAG/JOAG? ● Yes (Williams-Lyn et al., 2000) ● No (Faucher et al., 2002; Williams-Lyn et al., 2000) gnomAD? ● Allele count 337, allele frequency 1.19e-3 ○ African/African American	Secretion Assay ● 37 °C ○ +++ (Gobeil et al., 2006) ● 30 °C ○ n/a Triton X-100 Solubility Assay ● Soluble (Z. Zhou & Vollrath, 1999) ● Insoluble (Shimizu et al., 2000)	● WT-like based on Glu352Gln (Burns, Turnage, Walker, & Lieberman, 2011)	Location ● Surface exposed ● Exterior loop of Blade B Interactions ● None Predicted consequence of substitution ● Tolerated	n/a	n/a
p.Thr353Ile c.1058C>T	Diagnosis ● JOAG (Bhattacharjee et al., 2007; Lam et al., 2000; Park et al., 2016; Rose et al., 2011) ● POAG (Banerjee, Bhattacharjee, Ponda, Sen, & Ray, 2012; B. Fan et al., 2002; B. J. Fan et al., 2005; Fingert et al., 1999; Jia, Tam, et al., 2009; Lam et al., 2000; Pang et al., 2002; Yoon, Kim, Moon, Lim, & Joo, 1999; X. M. Zhou et al., 2013) ● NTG (B. J. Fan et al., 2005) ● PACG (Aung et al., 2005) ● Control Group(Aung et al., 2005; B. Fan et al., 2002; B. J. Fan et al., 2005; Lam et al., 2000; Liu et al., 2012; Pang et al., 2002) Country/Ethnicity ● Korean ● Japanese ● Chinese ● Indian Family History of POAG/JOAG? ● Yes (B. Fan et al., 2002; Park et al., 2016; Rose et al., 2011; Yoon et al., 1999; X. M. Zhou et al., 2013) ● No (Bhattacharjee et al., 2007) gnomAD? ● Allele count 191, allele frequency 6.75e-4 ○ Asian	Secretion Assay ● 37 °C ○ +++ (Gobeil et al., 2006) ● 30 °C ○ n/a Triton X-100 Solubility Assay ○ n/a	WT-like ○ T_m=53.1 °C (Donegan et al., 2015)	Location ● Surface exposed, outer strand of Blade B Interactions ● Thr351 side chain, solvent Predicted consequence of substitution ● Tolerated	n/a	● Homozygous carrier had more severe symptoms than heterozygous family members (Yoon et al., 1999) ● Individual heterozygous for Thr353Ile and Asp384Asn had more severe symptoms than father with only Thr353Ile (X. M. Zhou et al., 2013) ● One individual heterozygous for Thr353Ile and Asn480Lys (Rose et al., 2011)
p.Lys398Arg c.1193G>A	Diagnosis ● POAG (Alward et al., 1998; Alward et al., 2002; Fingert et al., 1999; Hulsman et al., 2002; Jansson, Marknell, Tomic, Larsson, & Wadelius, 2003; Liu et al., 2012; McDonald et al., 2010; O’Gorman et al., 2019; Rakhmanov et al., 2005; Vázquez, Herrero, Bastús, & Pérez, 2009; Vincent et al., 2002; Williams-Lyn et al., 2000) ● Control Group (Alward et al., 1998; Fingert et al., 1999; Hulsman et al., 2002; Liu et al., 2012; Vázquez et al., 2009) ● Exfoliative glaucoma (Jansson et al., 2003) ● Pigmentary glaucoma (Alward et al., 1998; Alward et al., 2002; Fingert et al., 1999) Country/Ethnicity ● Russian ● Mexican ● Dutch ● Swedish ● English/Scottish ● Canadian ● Spanish ● UK ○ African American ● Australian ● Caucasian Family History of POAG/JOAG? ○ n/a gnomAD? ● Allele count 992, allele frequency 3.51e-3 ● European	Secretion Assay ● 37 °C ○ +++ (Gobeil et al., 2006; Jacobson et al., 2001) ● 30 °C ○ n/a Triton X-100 Solubility Assay ● Soluble (Shimizu et al., 2000; Z. Zhou & Vollrath, 1999)	WT-like ○ T_m=53.8 °C (Burns et al., 2011)	Location ● Surface exposed, loop connecting inner strands of Blade C Interactions ● None Predicted consequence of substitution ● Tolerated	n/a	n/a
p.Ala445Val c.1334C>T	Diagnosis ● POAG (Alward et al., 1998; Faucher et al., 2002; Fingert et al., 1999; Lopez-Martinez et al., 2007) ● NTG (Weisschuh et al., 2005) ● Pigmentary glaucoma (Faucher et al., 2002) ● OHT (Vincent et al., 2002) Country/Ethnicity ● German ● Canadian ● French-Canadian ● Spanish ● Australian Family History of POAG/JOAG? ● Yes (Vincent et al., 2002) ● No (Faucher et al., 2002) gnomAD? ● Allele count 57, allele frequency 2.01e-4	Secretion Assay ● 37 °C ○ +++ (Gobeil et al., 2006) ● 30 °C ○ n/a Triton X-100 Solubility Assay ○ n/a	● WT-like ○ T_m=54.2 °C (Burns et al., 2011)	Location ● Surface exposed, on loop connecting inner strands of Blade D Interactions ● Thr443 Predicted consequence of substitution ● Tolerated	n/a	n/a
p.Lys500Arg c.1499A>G	Diagnosis ● POAG (Alward et al., 1998; Fingert et al., 1999; Liu et al., 2012; Whigham et al., 2011; Williams, Carmichael, Wainstein, Hobbs, & Ramsay, 2015; Williams-Lyn et al., 2000) ● Control (Liu et al., 2012) Country/Ethnicity ○ African American ● Black South African Family History of POAG/JOAG? ● Yes (Williams et al., 2015) gnomAD? ● Allele count 161, allele frequency 5.69e-4	Secretion Assay ● 37 °C ○ n/a ● 30 °C ○ n/a Triton X-100 Solubility Assay ● n/a	n/a	Location ● C-terminal strand within molecular clasp of Blade E, surface exposed Interactions ● None Predicted consequence of substitution ● Tolerated	p.Lys500Thr (c.1499A>C) ● gnomAD: Allele count 1, allele frequency 3.98e-6 ● Tolerated in structure	One control individual was compound heterozygous for p.Lys500Arg and p.Asp446Tyr (Liu et al., 2012)

B. Pathogenic
Variant	Clinical Data	Cell Assays	Thermal Stability	Structure Analysis	Additional Variants	Other

p.Cys245Tyr c.734G>A	Diagnosis ● JOAG (B. J. Fan et al., 2006) ● OHT (B. J. Fan et al., 2006) Country/Ethnicity ● Chinese Family History of POAG/JOAG? ● Yes (B. J. Fan et al., 2006) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ - (Hogewind et al., 2007) ○ 0.28% of WT (Zadoo, Nguyen, Zode, & Hulleman, 2016) ● 30 °C ○ −/+ (Zadoo et al., 2016) Triton X-100 Solubility Assay ● Insoluble (Hogewind et al., 2007; Yam, Gaplovska-Kysela, Zuber, & Roth, 2007) ● Partially insoluble (Jia, Gong, et al., 2009)	Likely destabilized ○ n/a (but see Cys433Arg)	Location ● Blade E, molecular clasp, surface exposed Interactions ● Disulfide bonded to Cys433 Predicted consequence of substitution ● Deleterious	n/a	n/a
p.Gly246Arg c.736G>A	Diagnosis ● JOAG (Adam et al., 1997; Brézin et al., 1997) Country/Ethnicity ● French Family History of POAG/JOAG? ● Yes (Adam et al., 1997; Brézin et al., 1997) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ - (Gobeil et al., 2006; Vollrath & Liu, 2006) ○ 0.47% of WT (Zadoo et al., 2016) ● 30 °C ○ ++ (Gobeil et al., 2006; Vollrath & Liu, 2006; Zadoo et al., 2016) Triton X-100 Solubility Assay ○ n/a	Destabilized ○ T_m=~42.6 °C (Burns et al., 2011; Donegan, Hill, Turnage, Orwig, & Lieberman, 2012)	Location ● Blade E, molecular clasp, surface exposed Interactions ● None Predicted consequence of substitution ● Deleterious, charged side chain like Arg would interfere with proper positioning of Cys245 for disulfide bond formation	n/a	n/a
p.Gly252Arg c.754G>A	Diagnosis ● POAG (Hewitt et al., 2007) ● JOAG (Booth et al., 2000; Rozsa et al., 1998; Shimizu et al., 2000; Vincent et al., 2002; Willoughby et al., 2004) ● OHT (Hewitt et al., 2007) Country/Ethnicity ● Australian ● Chinese ● Scottish ● US/Caucasian Family History of POAG/JOAG? ● Yes (Booth et al., 2000; Hewitt et al., 2007; Rozsa et al., 1998; Shimizu et al., 2000; Vincent et al., 2002) gnomAD? ○ n/a	Secretion Assay ● 37 °C ● – (Gobeil et al., 2006) ● 30 °C ○ +++ (Gobeil et al., 2006) Triton X-100 Solubility Assay ● Insoluble(Shimizu et al., 2000)	Destabilized ○ T_m=44.8 °C (Burns et al., 2011) or 43.0 °C (Donegan et al., 2012), buffer dependent	Location ● Blade E, molecular clasp, surface exposed Interactions ● Main chain NH H-bonded to carbonyl of Asp498 Predicted consequence of substitution ● Deleterious, Arg would clash with neighboring Glu253 and Trp250, causing destabilization of the molecular clasp	● p.Gly252Glu (c.755G>A) ○ gnomAD: allele count 2, allele frequency 8.25e-6 ○ Likely deleterious to structure ● p.Gly252Ala (c.755G>C) ○ gnomAD: allele count 3, allele frequency 1.15e-5 ○ Likely deleterious to structure	● One individual compound heterozygous for Gly252Arg and Gly244Val (Hewitt et al., 2007) ● One individual also heterozygous for OPTN mutation (Willoughby et al., 2004)
p.Arg272Gly c.814C>G	Diagnosis ● JOAG (Shimizu et al., 2000) ● POAG (Shimizu et al., 2000) Country/Ethnicity ● US/Caucasian Family History of POAG/JOAG? ● Yes (Shimizu et al., 2000) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ - (Gobeil et al., 2006) ● 30 °C ○ ++ (Vollrath & Liu, 2006) Triton X-100 Solubility Assay ● Insoluble (Shimizu et al., 2000)	Destabilized ○ T_m=44.7 °C (Burns et al., 2011) or 41 °C (Donegan et al., 2012), buffer dependent	Location ● Blade A, innermost β-strand, near bottom face, not solvent exposed Interactions ● Sidechain guanidinium group forms multiple stabilizing interactions with Glu483 on Blade E, and Thr285 on the adjacent β-strand of Blade A Predicted consequence of substitution ● Deleterious, removal of side chain interactions	● p.Arg272Gln (c.815G>A) ○ gnomAD: allele count 2, allele frequency 8.01e-6 ○ Likely deleterious to structure ● p.Arg272* (c.814C>T) ○ gnomAD: allele count 2, allele frequency 8.02e-6 ○ Uncertain	n/a
p.Glu323Lys c.967G>A	Diagnosis ● JOAG (Rozsa et al., 1998; Shimizu et al., 2000) ● POAG (Shimizu et al., 2000) Country/Ethnicity ● Panamanian Family History of POAG/JOAG? ● Yes (Rozsa et al., 1998; Shimizu et al., 2000) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ - (Gobeil et al., 2006)/+(Vollrath & Liu, 2006) ● 30 °C ○ +++ (Gobeil et al., 2006; Vollrath & Liu, 2006) Triton X-100 Solubility Assay ● Insoluble (Shimizu et al., 2000; Z. Zhou & Vollrath, 1999)	Destabilized ○ T_m=44 °C (Donegan et al., 2012) or 45.6 °C (Burns et al., 2011), buffer dependent	Location ● Loop connecting Blade A to Blade B, top face, but not surface exposed Interactions ● Main chain NH forms H-bonding interaction with carbonyl of Gln337 ● Side chain forms polar contact with NH Glu340 Predicted consequence of substitution ● Deleterious, charge inversion	● p.Glu323_Val329dup (c.967_987dup) ○ gnomAD: allele count 1, allele frequency 3.98e-6 ○ Likely deleterious to structure	n/a
p.Gly364Val c.1091G>T	Diagnosis ● POAG (Alward et al., 1998; Fingert et al., 1999; Stone et al., 1997) Country/Ethnicity ● US ● US/Caucasian Family History of POAG/JOAG? ● Yes(Stone et al., 1997) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ - (Gobeil et al., 2006)/++ (Jacobson et al., 2001; Vollrath & Liu, 2006) ● 30 °C ○ +++ (Gobeil et al., 2006; Jacobson et al., 2001; Vollrath & Liu, 2006) Triton X-100 Solubility Assay ● Partially insoluble (Z. Zhou & Vollrath, 1999)	Destabilized ○ T_m=~45 °C (Burns et al., 2011; Donegan et al., 2012)	Location ● Long loop connecting Blade B to Blade C, surface exposed Interactions ● Main chain NH forms H-bonding interaction with carbonyl of Ser393 Predicted consequence of substitution ● Deleterious, steric clash with Gln368 and Phe369	● p.Gly364Ser (c.1090G>A) ○ gnomAD: allele count 2, allele frequency 7.95e-6 ● Likely deleterious to structure	● Adenovirus-injected myocilin^G364V into mouse eyes led to significant IOP elevation (Shepard et al., 2007)
p.Gly367Arg c.1099G>A	Diagnosis ● JOAG (Cobb et al., 2002; Gupta et al., 2020; Iliev et al., 2008; Kanagavalli, Krishnadas, Pandaranayaka, Krishnaswamy, & Sundaresan, 2003; Souzeau et al., 2015; Vincent et al., 2002; Yao et al., 2018) ● POAG (J. Chen et al., 2011; Faucher et al., 2002; Mansergh et al., 1998; Melki, Belmouden, Brezin, & Garchon, 2003; K. G. Michels-Rautenstrauss et al., 1998; Suzuki et al., 1997; Fumiko Taniguchi, Suzuki, Shirato, & Araie, 2000) Country/Ethnicity ● Japanese ● German ● Irish ● Indian ● Swiss ● Australian ● Scottish ● Chinese ● French ● Canadian ● Italian/French Family History of POAG/JOAG? ● Yes (J. Chen et al., 2011; Cobb et al., 2002; Faucher et al., 2002; Gupta et al., 2020; Iliev et al., 2008; Mansergh et al., 1998; K. G. Michels-Rautenstrauss et al., 1998; Souzeau et al., 2015; Fumiko Taniguchi et al., 2000; Vincent et al., 2002; Yao et al., 2018) ● No (Gupta et al., 2020; Kanagavalli et al., 2003) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ - (Gobeil et al., 2006; Kanagavalli, Pandaranayaka, Krishnadas, Krishnaswamy, & Sundaresan, 2007) ● 30 °C ○ +++ (Gobeil et al., 2006) Triton X-100 Solubility Assay ○ n/a	Destabilized ○ T_m= 42.7 °C (Donegan et al., 2012) or 45.7 °C (Burns et al., 2011), buffer dependent	Location ● Long loop connecting Blade B to Blade C, surface exposed Interactions ● Main chain NH forms polar contact with side chain of Asp378 Predicted consequence of substitution ● Deleterious, large size of arginine would be sterically incompatible with neighboring Tyr376, which forms a solvent-exposed π-π stacking interaction with His366	See p.Gly367_Gln368delinsVal	n/a
p.Gly367_Gln368delinsVal c.1101_1103delGACAinsT	Diagnosis ● JOAG (Angius, De Gioia, et al., 1998; Angius, Pisano, et al., 1998) ● POAG (Angius, De Gioia, et al., 1998; Angius, Pisano, et al., 1998) Country/Ethnicity ● Italian Family History of POAG/JOAG? ● Yes (Angius, De Gioia, et al., 1998; Angius, Pisano, et al., 1998) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ - (Vollrath & Liu, 2006) ● 30 °C ● + (Vollrath & Liu, 2006) Triton X-100 Solubility Assay ● Insoluble (Z. Zhou & Vollrath, 1999)	n/a	Location ● Long loop connecting Blade B to Blade C, surface exposed Interactions ● Main chain NH forms polar contact with side chain of Asp378 Predicted consequence of substitution ● Deleterious, changes to this loop are generally problematic	See p.Gly367Arg	n/a
p.Pro370Leu c.1109C>T	Diagnosis ● JOAG (Adam et al., 1997; Bayat et al., 2008; Campos-Mollo et al., 2007; X. Chen et al., 2009; Y. Chen, Jiang, Wan, Yu, & Sun, 2006; Damji et al., 1999; Hamanaka et al., 2017; C. Huang et al., 2018; X. Huang et al., 2014; Li et al., 2014; Mukhopadhyay et al., 2002; Rose et al., 2011; Rozsa et al., 1998; Shimizu et al., 2000; Stoilova et al., 1998; Suzuki et al., 1997; Svidnicki et al., 2018; F. Taniguchi, 1999; Vasconcellos et al., 2000; Vincent et al., 2002; Wei et al., 2011; Wiggs et al., 1998) ● POAG (Wei et al., 2005) Country/Ethnicity ● Colombian ● Chinese ● Iranian ● Japanese ● French-Canadian ● Canadian ● Indian ● Brazilian ● Greek ● French ● US ● US/Caucasian Family History of POAG/JOAG? ● Yes (Adam et al., 1997; Campos-Mollo et al., 2007; X. Chen et al., 2009; Y. Chen et al., 2006; Damji et al., 1999; Hamanaka et al., 2017; X. Huang et al., 2014; Li et al., 2014; Mukhopadhyay et al., 2002; Rozsa et al., 1998; Shimizu et al., 2000; Stoilova et al., 1998; Suzuki et al., 1997; F. Taniguchi, 1999; Vincent et al., 2002; Wei et al., 2005; Wei et al., 2011; Wiggs et al., 1998) ● No (Bayat et al., 2008) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ - (Gobeil et al., 2006; Vollrath & Liu, 2006; Yam et al., 2007) ● 30 °C ○ - (Gobeil et al., 2006; Vollrath & Liu, 2006) Triton X-100 Solubility Assay ● Insoluble (Shimizu et al., 2000; Yam et al., 2007; Z. Zhou & Vollrath, 1999)	Destabilized ● Could not be isolated in folded state for measurement (Burns et al., 2011) ● When combined with stabilizing variant D478S to yield OLF^P370L/D478S, T_m = 47°C compared to OLF^D478S alone, which is 58 °C (Hill et al., 2019)	Location ● Long loop connecting Blade B to Blade C, surface exposed Interactions ● None Predicted consequence of substitution ● Deleterious, adjacent to Tyr371, likely Pro likely properly organizes this region to promote the interaction between the backbone carbonyl of Phe369 and Nε of Lys423, and orient Tyr371 for cation-π	● p.Pro370Ser (c.1108C>T) ○ gnomAD: allele count 1, allele frequency 3.19e-5 ○ Likely deleterious to structure	● Often serves as a “positive control” for pathogenic variants ● Myocilin^P370L accumulates in the ER(Lopez-Martinez et al., 2007) ● Myocilin^P370L downregulates homeostatic ER-stress response (L. Wang et al., 2007) ● Myocilin^P370L impairs mitochondrial function (decreased transmembrane potential, increased ROS production, increased intracellular cytoplasm) (He, Leung, Zhuo, & Ge, 2009) ● Detailed histochemical analysis (Hamanaka et al., 2017)
p.Thr377Met c.1130C>T	Diagnosis ● JOAG (Puska, Lemmela, Kristo, Sankila, & Jarvela, 2005; Shimizu et al., 2000; Young et al., 2012; Zgaga et al., 2008) ● POAG (Allingham et al., 1998; Alward et al., 1998; Fingert et al., 1999; Hamanaka et al., 2017; Kanagavalli et al., 2003; Kitsos et al., 2010; Liu et al., 2012; Mackey et al., 2003; Melki, Idhajji, et al., 2003; Petersen et al., 2006; Puska et al., 2005; Shimizu et al., 2000; Wiggs et al., 1998; Wilkinson et al., 2003; Wirtz et al., 2008) ● NTG (Mackey et al., 2003; Puska et al., 2005; Wilkinson et al., 2003) ● OHT (Hamanaka et al., 2017; Kitsos et al., 2010; Mackey et al., 2003; Petersen et al., 2006; Puska et al., 2005; Wilkinson et al., 2003; Wirtz et al., 2008) Country/Ethnicity ● Croatian ● US ● US/Caucasian ● Indian ● Australian ○ African American ● Finnish ● Greek ● Dutch/Croatian ● English ● Greek/Macedonian ● Japanese ● Moroccan Family History of POAG/JOAG? ● Yes(Allingham et al., 1998; Alward et al., 1998; Hamanaka et al., 2017; Kanagavalli et al., 2003; Kitsos et al., 2010; Mackey et al., 2003; Petersen et al., 2006; Puska et al., 2005; Shimizu et al., 2000; Wiggs et al., 1998; Wilkinson et al., 2003; Wirtz et al., 2008; Young et al., 2012; Zgaga et al., 2008) gnomAD? ● Allele count 5, allele frequency 1.99e-5	Secretion Assay ● 37 °C ● + (Gobeil et al., 2006)/++ (Vollrath & Liu, 2006) ● 30 °C ○ +++ (Gobeil et al., 2006; Vollrath & Liu, 2006) Triton X-100 Solubility Assay ● Partially insoluble (Z. Zhou & Vollrath, 1999) ● Insoluble (Shimizu et al., 2000)	Destabilized ● Tm = 44.3 °C (Donegan et al., 2012) or 47.7°C (Burns et al., 2011), buffer dependent	Location ● Long loop connecting Blade B to Blade C Interactions ● Main chain carbonyl makes polar contact with Nε of Lys423 ● Side chain makes polar contacts with Gly374 carbonyl and main chain NH of Tyr371 Predicted consequence of substitution ● Deleterious, loss of numerous polar interactions would propagate to weaken cation-π and metal ion site	● p.Thr377Lys (c.1130C>A) (Vincent et al., 2002) ○ Likely deleterious to structure ● p.Thr377Arg (c.1130C>G) (Waryah, Narsani, Sheikh, Shaikh, & Shahani, 2013) ○ Likely deleterious to structure ● p.Thr377Ala (c.1129A>G) ○ gnomAD: allele count 2, allele frequency 7.96e-6 ○ Uncertain	● Detailed histochemical analysis (Hamanaka et al., 2017) ● Four family members compound heterozygous for p.Thr377Met and p.Gln368* have higher IOP than family members with only p.Gln368* (Young et al., 2012) ● Two family members compound heterozygotes for p.Thr377Met and p.Val495Ile (Alward et al., 1998)
p.Asp380Ala c.1139A>C	Diagnosis ● JOAG (Campos-Mollo et al., 2007; Kennan et al., 1998; Stoilova et al., 1998) Country/Ethnicity ● Spanish ● English Family History of POAG/JOAG? ● Yes (Campos-Mollo et al., 2007; Kennan et al., 1998; Stoilova et al., 1998) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ - (Gobeil et al., 2006)/+(Vollrath & Liu, 2006) ● 30 °C ○ ++ (Gobeil et al., 2006)/+++ (Vollrath & Liu, 2006) Triton X-100 Solubility Assay ● Partially insoluble (Z. Zhou & Vollrath, 1999)	Destabilized ● Tm = ~46.6 °C (Burns et al., 2011; Donegan et al., 2012)	Location ● Innermost strand, Blade C Interactions ● Ca²⁺, Na⁺, Tyr371 Predicted consequence of substitution ● Deleterious, loss of ionic interactions	● p.Asp380Gly (c.1139A>G) (Alward et al., 1998) ○ Likely deleterious to structure ● p.Asp380His (c.1138G>C) (Wirtz, Samples, Choi, & Gaudette, 2007) ○ Likely deleterious to structure ● p.Asp380Asn (c.1138G>A) (Challa et al., 2002) ○ Likely deleterious to structure	● Aggregation of OLF^D380A is facile at 37 °C, pH 7.2 (Hill, Donegan, & Lieberman, 2014)
p.Lys423Glu c.1267A>G	Diagnosis ● JOAG (Bruttini et al., 2003; Faucher et al., 2002; Morissette et al., 1998; Svidnicki et al., 2018) ● POAG (Bruttini et al., 2003; Morissette et al., 1998) ● OHT (Morissette et al., 1998) Country/Ethnicity ● Brazilian ● Italian ● French-Canadian ● Canadian Family History of POAG/JOAG? ● Yes (Bruttini et al., 2003; Faucher et al., 2002; Morissette et al., 1998) ● No (Faucher et al., 2002) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ - (Gobeil et al., 2006; Jacobson et al., 2001; Vollrath & Liu, 2006) ● 30 °C ○ - (Gobeil et al., 2006; Vollrath & Liu, 2006) Triton X-100 Solubility Assay ● Insoluble (Z. Zhou & Vollrath, 1999)	Destabilized ● 34.2 °C (Donegan et al., 2012) or 40.5 °C (Burns et al., 2011)	Location ● On loop connecting Blade C to Blade D Interactions ● Internal cation-π with Tyr371 Predicted consequence of substitution ● Deleterious, charge inversion internal to protein	n/a	● In family, 4 homozygotes had no symptoms while heterozygotes did have JOAG/POAG (Morissette et al., 1998)
p.Val426Phe c.1276G>T	Diagnosis ● JOAG (Lim, Lichter, Higashi, Downs, & Richards, 2003; Mansergh et al., 1998; Millá et al., 2013; Rozsa et al., 1998; Shimizu et al., 2000) ● POAG (Shimizu et al., 2000) Country/Ethnicity ● Spanish ● US/Caucasian Family History of POAG/JOAG? ● Yes (Lim et al., 2003; Mansergh et al., 1998; Millá et al., 2013; Rozsa et al., 1998; Shimizu et al., 2000) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ - (Gobeil et al., 2006; Vollrath & Liu, 2006) ● 30 °C ○ ++ (Vollrath & Liu, 2006)/+++ (Gobeil et al., 2006) Triton X-100 Solubility Assay ● Insoluble (Shimizu et al., 2000; Z. Zhou & Vollrath, 1999)	Destabilized ○ T_m= 41.5 °C (Donegan et al., 2012) or 45.1 °C (Burns et al., 2011), buffer dependent	Location ● On loop connecting Blade C to Blade D Interactions ● Hydrophobic between Blades C and D Predicted consequence of substitution ● Deleterious, would weaken or disrupt the neighboring cation-π interaction between Lys423 and Tyr371 (see Lys423)	n/a	n/a
p.Cys433Arg c.1297T>C	Diagnosis ● JOAG (Povoa, Malta, Rezende Mde, de Melo, & Giannella-Neto, 2006; Svidnicki et al., 2018; Vasconcellos et al., 2000) ● POAG (Povoa et al., 2006) ● OHT (Povoa et al., 2006) Country/Ethnicity ● Brazilian Family History of POAG/JOAG? ● Yes (Povoa et al., 2006; Vasconcellos et al., 2000) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ - (Gobeil et al., 2006) ● 30 °C ○ - (Gobeil et al., 2006) Triton X-100 Solubility Assay ○ n/a	Destabilized ○ T_m=40.4 °C (Burns et al., 2011) or 42.6 °C (Donegan et al., 2012), buffer dependent	Location ● Blade D, β-turn connecting two inner strands on bottom face Interactions ● Disulfide with Cys245, and NH fmors interaction with Glu385; Glu385Ala is misfolded (Donegan et al., 2012) Predicted consequence of substitution ● Deleterious, loss of covalent disulfide bond and interacting with Glu385	n/a	n/a
p.Tyr437His c.1309T>C	Diagnosis ● JOAG (Alward et al., 2002; Lei, Li, Liu, & Zhang, 2019; Wiggs et al., 1998; Yang et al., 2020) ● POAG (Alward et al., 1998; Alward et al., 2002; Fingert et al., 1999; Lei et al., 2019; Stone et al., 1997) ● OHT (Lei et al., 2019) Country/Ethnicity ● Chinese ● US ● US/Caucasian Family History of POAG/JOAG? ● Yes (Alward et al., 1998; Lei et al., 2019; Stone et al., 1997; Wiggs et al., 1998; Yang et al., 2020) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ - (Gobeil et al., 2006; Jacobson et al., 2001; Vollrath & Liu, 2006) ○ 0.2% of WT (Zadoo et al., 2016) ● 30 °C ○ −/+ (Vollrath & Liu, 2006; Zadoo et al., 2016) Triton X-100 Solubility Assay ● Insoluble (Hogewind et al., 2007; Z. Zhou & Vollrath, 1999)	Destabilized ○ T_m=40.3 °C (Donegan et al., 2012) or 41.4 °C (Burns et al., 2011), buffer dependent	Location ● Blade D, near disulfide bond Interactions ● Side chain forms H-bonding interaction with NH of Leu248 in Blade E Predicted consequence of substitution ● Deleterious, His is too small to form inter-blade interaction	n/a	● Detailed histochemical comparison between human donor eyes expressing myocilin^Y437H or myocilin^WT (van der Heide et al., 2018) ● Mouse models with different genetic backgrounds have differing phenotypes (Chou, Tomarev, & Porciatti, 2014; Gould et al., 2006; Joe, Nakaya, Abu-Asab, & Tomarev, 2015; Kasetti, Phan, Millar, & Zode, 2016; Lynch et al., 2018; McDowell et al., 2012; Senatorov et al., 2006; Shepard et al., 2007; Zhuo et al., 2008; Zillig, Wurm, Grehn, Russell, & Tamm, 2005; Zode et al., 2011) ● Aggregated OLF^Y437H has amyloid character (Orwig et al., 2012)
p.Ile477Asn c.1430T>A	Diagnosis ● JOAG (Richards, 1998; Rozsa et al., 1998; Shimizu et al., 2000) ● POAG (Alward et al., 1998; Fingert et al., 1999; Shimizu et al., 2000) ● Country/Ethnicity ● US ● US/Caucasian Family History of POAG/JOAG? ● Yes (Alward et al., 1998; Richards, 1998; Rozsa et al., 1998; Shimizu et al., 2000; Stokes, 1940) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ - (Gobeil et al., 2006; Vollrath & Liu, 2006) ● 30 °C ○ - (Gobeil et al., 2006; Vollrath & Liu, 2006) Triton X-100 Solubility Assay ● Insoluble (Shimizu et al., 2000; Z. Zhou & Vollrath, 1999)	Destabilized ○ T_m=37.7 °C (Burns et al., 2011) or 39.7 °C (Donegan et al., 2012), buffer dependent	Location ● Blade E, innermost strand, part of hydrophobic belt Interactions ● Side chain hydrophobic packing, main chain carbonyl is ligand to Ca²⁺ Predicted consequence of substitution ● Deleterious, new polarity in hydrophobic environment	See p.Ile477Ser	n/a
p.Ile477Ser c.1430T>G	Diagnosis ● POAG (Adam et al., 1997) Country/Ethnicity ● French Family History of POAG/JOAG? ● Yes (Adam et al., 1997) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ - (Izumi et al., 2003; Jacobson et al., 2001; Vollrath & Liu, 2006) ● 30 °C ○ +++ (Vollrath & Liu, 2006) Triton X-100 Solubility Assay ● Insoluble (Z. Zhou & Vollrath, 1999)	Destabilized ○ T_m = 40.1 °C (Donegan et al., 2012) or 41.9 °C (Burns et al., 2011), buffer dependent	Location ● Blade E, innermost strand, part of hydrophobic belt Interactions ● Side chain hydrophobic packing, main chain carbonyl is ligand to Ca²⁺ Predicted consequence of substitution ● Deleterious, new polarity in hydrophobic environment	See p.Ile477Asn	n/a
p.Tyr479His c.1435T>C	Diagnosis ● POAG (Lopez-Martinez et al., 2007; Millá et al., 2013) Country/Ethnicity ● Spanish Family History of POAG/JOAG? ● Yes (Millá et al., 2013) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ −/+ (Lopez-Martinez et al., 2007) ● 30 °C ○ n/a Triton X-100 Solubility Assay ○ n/a (Z. Zhou & Vollrath, 1999)	n/a	Location ● Blade E, innermost strand, near bottom face Interactions ● Tyr479 side chain forms short (2.6 Å) H-bonding interaction with Glu385, located on Blade C, otherwise in hydrophobic environment Predicted consequence of substitution ● Deleterious, His is too small to form interaction with Glu385, and Glu385Ala is misfolded (Donegan et al., 2012), indicating the H-bonding interaction is important.	n/a	● Myocilin^Y479H accumulates in ER (Lopez-Martinez et al., 2007)
p.Asn480Lys c.1440C>A,G	Diagnosis ● JOAG (Brézin et al., 1998; Mimivati, Nurliza, Marini, & Liza-Sharmini, 2014; Rose et al., 2011) ● POAG (Adam et al., 1997; Brézin et al., 1998; Guevara-Fujita et al., 2008; Hulsman et al., 2002; Melki, Belmouden, et al., 2003) Country/Ethnicity ● Andean ● Malay ● French ● Dutch ● Indian Family History of POAG/JOAG? ● Yes (Adam et al., 1997; Brézin et al., 1998; Guevara-Fujita et al., 2008; Hulsman et al., 2002; Mimivati et al., 2014; Rose et al., 2011) gnomAD? ○ n/a	Secretion Assay ● 37 °C ● – (Gobeil et al., 2006)/+(Vollrath & Liu, 2006) ● 30 °C ● + (Gobeil et al., 2006)/++(Vollrath & Liu, 2006) Triton X-100 Solubility Assay ● Insoluble (Z. Zhou & Vollrath, 1999)	Destabilized ○ T_m=42.4 °C (Donegan et al., 2012) or 46.1 °C (Burns et al., 2011), buffer dependent	Location ● Blade E, on β-turn between internal strands, side chain buried Interactions ● Asn480 side chain forms polar interactions with main chain NH of Leu483 and Glu483 Predicted consequence of substitution ● Deleterious, buried point charge unfavorable	n/a	● One individual is compound heterozygous for Asn480Lys and Thr353Ile (Rose et al., 2011)
p.Pro481Thr c.1441C>A	Diagnosis ● POAG (Fingert et al., 1999) ● JOAG (Alward et al., 2002) Country/Ethnicity ● US Family History of POAG/JOAG? ○ n/a gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ n/a ● 30 °C ○ n/a Triton X-100 Solubility Assay ○ n/a		Location ● Blade E, on β-turn between internal strands, side chain buried Interactions ● Pro481 forms H-bonding interaction to Lys484 Nε in same loop Predicted consequence of substitution ● Deleterious, removal of constraint within β-turn	● See p.Pro481Leu ● p.Pro481Arg (c.1442C>G) (Jansson et al., 2003) ● Likely deleterious to structure ● p.Pro481Ser (c.1441C>T) (Mabuchi et al., 2001) ● gnomAD: allele count 1, frequency 3.98e-6 ● Uncertain	n/a
p.Pro481Leu c.1442C>T	Diagnosis ● POAG (Faucher et al., 2002; Fingert et al., 1999) ● OHT (Faucher et al., 2002) ● Angle Closure Glaucoma (Faucher et al., 2002) Country/Ethnicity ○ African American ● Canadian Family History of POAG/JOAG? ● Ye s(Faucher et al., 2002) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ - (Gobeil et al., 2006) ● 30 °C ○ ++ (Gobeil et al., 2006) Triton X-100 Solubility Assay ○ n/a	Destabilized ○ T_m= 45.5 °C (Donegan et al., 2012) or 46.6 °C (Burns et al., 2011), buffer dependent	Location ● Blade E, on β-turn between internal strands, side chain buried Interactions ● Pro481 forms H-bonding interaction to Lys484 N in same loop Predicted consequence of substitution ● Deleterious, removal of constraint within β-turn	See p.Pro481Thr	n/a
p.Ile499Phe c.1495A>T	Diagnosis ● POAG (Adam et al., 1997; Melki, Belmouden, et al., 2003) Country/Ethnicity ● French Family History of POAG/JOAG? ● Yes (Adam et al., 1997) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ - (Gobeil et al., 2006)/+ (Vollrath & Liu, 2006) ● 30 °C ○ +++ (Gobeil et al., 2006; Vollrath & Liu, 2006) Triton X-100 Solubility Assay ● Partially Insoluble (Z. Zhou & Vollrath, 1999)	Destabilized ○ T_m =42.8°C (Donegan et al., 2012) or 44.4°C (Burns et al., 2011), buffer dependent	Location ● Blade E, exterior strand, side chain buried in hydrophobic belt Interactions ● Hydrophobic Predicted consequence of substitution ● Deleterious, side chain to large	See p.Ile499Ser	n/a
p.Ile499Ser c.1496T>G	Diagnosis ● JOAG (Shimizu et al., 2000) Country/Ethnicity ● US/Caucasian Family History of POAG/JOAG? ● Yes (Shimizu et al., 2000) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ n/a ● 30 °C ○ n/a Triton X-100 Solubility Assay ● Insoluble (Shimizu et al., 2000)	Likely destabilized ○ n/a (but see Ile499Phe)	Location ● Blade E, exterior strand, side chain buried in hydrophobic belt Interactions ● Hydrophobic Predicted consequence of substitution ● Deleterious, new polarity in hydrophobic environment	See p.Ile499Phe	n/a

p.Ser502Pro c.1504T>C	Diagnosis ● JOAG (Stoilova et al., 1998) Country/Ethnicity ● English Family History of POAG/JOAG? ● Yes (Stoilova et al., 1998) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ - (Gobeil et al., 2006) ● 30 °C ○ - (Gobeil et al., 2006) Triton X-100 Solubility Assay ○ n/a	Destabilized ○ T_m= 41.0 °C (Donegan et al., 2012) or 43.0 °C (Burns et al., 2011), buffer dependent	Location ● Blade E, molecular clasp, surface exposed Interactions ● Main chain to Glu247 Predicted consequence of substitution ● Deleterious, new backbone constraint	n/a	● Adenovirus-injected myocilin^S502P into mouse eyes did not result in IOP elevation (Shepard et al., 2007)

C. Likely Pathogenic
Variant	Clinical Data	Cell Assays	Thermal Stability	Structure Analysis	Additional Variants	Other

p.Val251Ala c.752T>C	Diagnosis ● JOAG (Kuchtey, Chowdhury, Uptegraft, Fautsch, & Kuchtey, 2013; Lu et al., 2020; K. Michels-Rautenstrauss et al., 2002) ● POAG (Lu et al., 2020) ● OHT (Lu et al., 2020) Country/Ethnicity ● German ● Chinese ● US/Caucasian Family History of POAG/JOAG? ● Yes (Kuchtey et al., 2013; Lu et al., 2020; K. Michels-Rautenstrauss et al., 2002) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ n/a ● 30 °C ○ n/a Triton X-100 Solubility Assay ○ n/a	n/a	Location ● Blade E, molecular clasp, side chain pointed inward toward hydrophobic belt Interactions ● None Predicted consequence of substitution ● Uncertain, Val to Ala substitution is conservative, but weaker hydrophobic packing	● p.Val251Ile (c.751G>A) ○ gnomAD: allele count 1, allele frequency 4.14e-6 ○ Likely deleterious to structure	● Myocilin^V251A was not found in the aqueous humor of a patient with JOAG (Kuchtey et al., 2013)
p.Pro254Arg c.761C>G	Diagnosis ● JOAG (Yang et al., 2015) ● POAG (Yang et al., 2015) Country/Ethnicity ● Chinese Family History of POAG/JOAG? ● Yes (Yang et al., 2015) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ n/a ● 30 °C ○ n/a Triton X-100 Solubility Assay ○ n/a	n/a	Location ● Blade E, molecular clasp Interactions ● Main chain carbonyl forms polar interactions with Arg470, located on an opposing loop that connects Blade D and Blade E Predicted consequence of substitution ● Deleterious, would remove backbone restriction in molecular clasp and side chain too large	● p.Pro254Leu (c.761C>T) (Souzeau et al., 2016) ○ Likely deleterious to structure	n/a
p.Leu255Pro c.764T>C	Diagnosis ● JOAG (Park et al., 2016) Country/Ethnicity ● Korean Family History of POAG/JOAG? ● Yes (Park et al., 2016) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ n/a ● 30 °C ○ n/a Triton X-100 Solubility Assay ○ n/a	n/a	Location ● Blade E, outer strand, surface exposed Interactions ● Main chain H-bonds with Thr496 main chain Predicted consequence of substitution ● Deleterious, would introduce di-proline motif, further limiting the polypeptide backbone mobility	● p.Leu255Val (c.763G>C) ○ gnomAD: allele count 3, allele frequency 1.23e-5 ○ Uncertain	n/a
p.Pro274Arg c.821C>G	Diagnosis ● JOAG (Markandaya et al., 2004) Country/Ethnicity ● Indian Family History of POAG/JOAG? ● Yes (Markandaya et al., 2004) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ n/a ● 30 °C ○ n/a Triton X-100 Solubility Assay ○ n/a	n/a	Location ● Blade A, loop connecting innermost strands Interactions ● None Predicted consequence of substitution ● Deleterious, Arg would remove restrictions placed on polypeptide backbone and also disrupt packing	● p.Pro274Leu (c.821C>T) ○ gnomAD: allele count 1, allele frequency 4e-6 ○ Likely deleterious to structure	● Youngest family member homozygous for mutation(Markandaya et al., 2004)
p.Gln337Arg ● c.1010A>G	Diagnosis ● JOAG (Gupta et al., 2020; Stoilova et al., 1997) Country/Ethnicity ● Scottish ● Indian Family History of POAG/JOAG? ● Yes (Gupta et al., 2020) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ n/a ● 30 °C ○ n/a Triton X-100 Solubility Assay ○ n/a	n/a	Location ● Blade B, loop connecting inner strands Interactions ● Main chain carbonyl forms polar contact with Glu323 main chain NH ● Side chain forms polar contact with main chain carbonyl of Ser341 Predicted consequence of substitution ● Deleterious, sterically incompatible	● See p.Gln337* (c.1009delC) ● p.Gln337Glu (c.1009C>G) (Vázquez et al., 2009) ○ Likely deleterious to structure ● p.Gln337His (c.1011G>T) ○ gnomAD: allele count 1, allele frequency 3.98e-6 ○ Likely deleterious to structure	● Homozygous individuals identified in two families (Gupta et al., 2020)
p.Ser341Pro c.1021T>C	Diagnosis ● JOAG (X. Huang et al., 2014) ● POAG (X. Huang et al., 2014; Kee & Ahn, 1997; Moon, Kim, & Lee, 2020; F. Wang et al., 2015) ● OHT (F. Wang et al., 2015) Country/Ethnicity ● Chinese ● Korean Family History of POAG/JOAG? ● Ye s(X. Huang et al., 2014; Kee & Ahn, 1997; Moon et al., 2020; F. Wang et al., 2015) ● No (X. Huang et al., 2014) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ n/a ● 30 °C ○ n/a Triton X-100 Solubility Assay ○ n/a	n/a	Location ● Blade B, turn between internal strands Interactions ● Main chain carbonyl forms polar contact with Gln337 side chain, main chain NH forms polar contact with carbonyl of Gly338 ● Side chain hydroxyl forms polar interactions with NH Thr343 and CO Gly338 Predicted consequence of substitution ● Deleterious, restrict backbone and reduce available stabilizing interactions	n/a	n/a
p.Ala363Thr c.1087G>A	Diagnosis ● POAG (Ishikawa et al., 2004; R. Kubota et al., 2000; Mengkegale et al., 2008) Country/Ethnicity ● Japanese Family History of POAG/JOAG? ● Yes (R. Kubota et al., 2000; Ryo Kubota et al., 1997; Mengkegale et al., 2008) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ - (Izumi et al., 2003) ● 30 °C ○ n/a Triton X-100 Solubility Assay ○ n/a	n/a	Location ● Long loop between Blade B and Blade C, pointed inward Interactions ● Main chain NH forms polar contact with carbonyl Ile360, main chain carbonyl forms polar contact with side chain Arg342 Predicted consequence of substitution ● Uncertain, relatively conservative substitution but introduces new polarity into relatively hydrophobic region	n/a	n/a
p.Phe369Leu c.1105T>C	Diagnosis ● JOAG (Hamanaka et al., 2017) ● POAG (Hamanaka et al., 2017; Ishikawa et al., 2004) Country/Ethnicity ● Japanese Family History of POAG/JOAG? ● Yes (Hamanaka et al., 2017; Ishikawa et al., 2004) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ n/a ● 30 °C ○ n/a Triton X-100 Solubility Assay ○ n/a	n/a	Location ● Long loop between Blade B and Blade C, surface exposed Interactions ● Main chain carbonyl makes polar contact with Lys423 Nε Predicted consequence of substitution ● Uncertain, Phe369 protects Lys423 from solvent, unclear if Leu would suffice	n/a	● One patient homozygous for variant (Hamanaka et al., 2017) ● Detailed histochemical analysis (Hamanaka et al., 2017)
p.Tyr371Asp c.1111T>G	Diagnosis ● JOAG (Avisar et al., 2009) Country/Ethnicity ● Uzbek Family History of POAG/JOAG? ● Yes (Avisar et al., 2009) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ n/a ● 30 °C ○ n/a Triton X-100 Solubility Assay ○ n/a	n/a	Location ● Long loop between Blade B and Blade C, points inward Interactions ● Main chain NH forms polar contact with side chain hydroxyl of Thr377; carbonyl forms polar contact with main chain NH of Trp373 ● Side chain ring forms cation-π with Lys423 and hydroxyl forms polar interaction with Asp380 Predicted consequence of substitution ● Deleterious, based on analogy with Lys423Glu	n/a	n/a
p.Glu385Lys c.1153G>A	Diagnosis ● JOAG (Criscione et al., 2019) Country/Ethnicity ● Hispanic Family History of POAG/JOAG? ● Ye s(Avisar et al., 2009; Criscione et al., 2019) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ n/a ● 30 °C ○ n/a Triton X-100 Solubility Assay ○ n/a	Likely destabilized ○ T_m= 39.7 °C for OLF^{E385A (Donegan et al., 2012)}	Location ● Blade C, bottom face, on turn between innermost blades Interactions ● Side chain forms polar contacts with main chain NH of Cys433 on Blade D, and with sidechains of Tyr 479 and Lys484 on Blade E Predicted consequence of substitution ● Deleterious, critical interactions would be lost	n/a	n/a
p.Thr438Ile c.1313C>T	Diagnosis ● JOAG (Jansson et al., 2003; Melki, Belmouden, et al., 2003; Souzeau et al., 2015) Country/Ethnicity ● Australian ● French ● Danish Family History of POAG/JOAG? ● Yes (Jansson et al., 2003; Melki, Belmouden, et al., 2003; Souzeau et al., 2015) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ n/a ● 30 °C ○ n/a Triton X-100 Solubility Assay ○ n/a	n/a	Location ● Blade D, interior strand Interactions ● Side chain hydroxyl group forms polar interaction with Asn450 sidechain Predicted consequence of substitution ● Deleterious, polar intra-blade contact would be lost	● p.Thr438Pro (c.1312A>C) ○ gnomAD: allele count 6, allele frequency 2.39e-5 ○ Likely deleterious to structure	n/a
p.Asn450Tyr c.1348A>T	Diagnosis ● JOAG (Zhao et al., 2010) ● OHT (Zhao et al., 2010) Country/Ethnicity ● Chinese Family History of POAG/JOAG? ● Yes (Zhao et al., 2010) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ n/a ● 30 °C ○ n/a Triton X-100 Solubility Assay ○ n/a	n/a	Location ● Blade D, interior strand, points to surface Interactions ● Side chain forms polar contact with Thr438 side chain hydroxyl Predicted consequence of substitution ● Deleterious, Tyr would pose a steric challenge with the loop connecting Blade C and Blade D	● p.Asn450Asp (c.1348A>G) (K. Michels-Rautenstrauss et al., 2002) ○ Likely tolerated in the structure ● p.Asn450Ser (c.1349A>G) ○ gnomAD: allele count 2, allele frequency 7.95e-6 ○ Likely tolerated in the structure	n/a
p.Leu486Phe ● c.1456C>T	Diagnosis ● JOAG (Liao et al., 2016) ● POAG (X. Huang et al., 2014) Country/Ethnicity ● Chinese Family History of POAG/JOAG? ● Ye s(Liao et al., 2016) ● No (X. Huang et al., 2014) gnomAD? ○ n/a	Secretion Assay ● 37 °C ○ n/a ● 30 °C ○ n/a Triton X-100 Solubility Assay ○ n/a	n/a	Location ● Blade E, inner strand, pointed inward Interactions ● Main chain NH forms polar interaction with Thr496 sidechain; main chain carbonyl forms polar contact with carbonyl of Tyr497 Predicted consequence of substitution ● Deleterious, would require rearrangements to fit larger side chain	n/a	n/a

qualitative extent of secretion − = no secretion, −/+ = very little secretion, +, ++, +++ increasing levels of secretion