Table 2.

Results gathered on pLG72 by employing deletion variants.

Variant	Region	Method	Properties				References
			Kd, hDAAO (μM)	Tm (°C)	Residual hDAAO activitya (%)	Comments
Wild-type (1–153)		RE	2.66	59.2	16	Mitochondrial; binds hDAAO with no change in kinetic parameters	Sacchi et al., 2008; Birolo et al., 2016
1–64		RE	2.2	73.7	72	Compared to the wild-type, the theoretical buried solvent accessible area following hDAAO binding is halved	Birolo et al., 2016
1–94			2.1	50.9	62
1–123			0.4	66.3	47
72–153			3.7	64.6	50
122–153		PS			110	Unspecific binding
26–153		RC				Partial loss of mitochondrial localization	Kvajo et al., 2008
36–153						Partial loss of mitochondrial localization
51–153
1–110						Low expression yield (probably because of protein instability)
1–120
1–20		Y2				No interaction with hDAAO	Chang et al., 2014
1–35
1–60
1–122
1–137
95–153						Interacts with hDAAO
123–153						Interacts with hDAAO; 1.4-fold more efficient in hDAAO coimmunoprecipitation than wild-type; increases hDAAO activity (20–30%)b
138–153						Interacts with hDAAO; 2.2-fold more efficient in hDAAO coimmunoprecipitation than wild-type; increases hDAAO activity
1–71		MM				Similar to membrane bound proteins (e.g., membrane transporters or O-GlcNAc transferase)	Kato and Fukui, 2017
72–153						Similar to adaptor proteins (type I DNA methyl transferase, RH bundle subdomain of G protein-coupled receptor kinase)

RE, characterization of pure recombinant protein expressed in E. coli; RC, overproduction in COS-7 cells; Y2, yeast two-hybrid system; PS, peptide synthesis; MM, molecular modeling.

^aAt 1:2 hDAAO:pLG72 ratio and 30 min of incubation.

^b3.5-fold increase in hDAAO activity with wild-type pLG72.