Table 1.
Properties of neuroligin-1 mutants
| Mutant name | Mutations (all in NL1 lacking inserts in splice sites A and B | Localization | α/β-Neurexin binding | Binding affinity (Kd) | Assays employed | Reference | |
|---|---|---|---|---|---|---|---|
| α | β | ||||||
| NL1 WT | Wild type | PM | + | + | 20.9 nM | ITC/SPR/SBA | Arac et al (2007) |
| NL1–1 | D242N/Q243A/K246A | ER | ND | ND | ND | SBA | This study |
| NL1–2 | Q256A/R259A | ER | ND | ND | ND | SBA | This study |
| NL1–3 | Q328A/Y332A | PM | + | + | ND | SBA | This study |
| NL1–4 | R371A/Y372A/H373A | PM | + | + | ND | SBA | This study |
| NL1–5 | L399A/N400A/D402N | PM | − | + | >10 μM | SBA | Arac et al (2007) |
| NL1–6 | D421N/D423N/D424N | PM | + | + | ND | SBA | This study |
| NL1–7 | D440N/N441A/D424N | PM | + | + | ND | SBA | This study |
| NL1–8 | N468A/E470A/R472A/K474A | ER | ND | ND | ND | SBA | This study |
| NL1–9 | E542Q/L543A/F544A | ER | ND | ND | ND | SBA | This study |
| NL1–10 | F582A/K586A/R589A | PM | + | + | ND | SBA | This study |
| NL1–11 | E128Q/V129A | PM | + | + | ND | SBA | This study |
| NL1–21 | E72Q/I73A | PM | + | + | ND | SBA | This study |
| NL1–22 | D140N | PM | + | + | ND | SBA | This study |
| NL1–23 | H294A/E297A | ER | ND | ND | ND | SBA | This study |
| NL1–24 | L302A/S321A/S322A | ER | ND | ND | ND | SBA | This study |
| NL1–25 | Y509A/H511A/Q513A | PM | + | + | ND | SBA | This study |
| NL1–31 | L399A/N400A/D402N/K306A | PM | − | + | ND | SBA | This study |
| NL1–32 | L399A/N400A/D402N/E297A/K306A | PM | − | − | >10 μM | SPR/SBA | Arac et al (2007) |
| NL1–33 | L399A/N400A/D402N/L273A | PM | − | + | ND. | SBA | This study |
| NL1–34 | L399A/N400A/D402N/K601A/D602A | PM | − | + | ND | SBA | This study |
| NL1–35 | L399A/N400A/D402N/Q395A/E397A | PM | − | − | >50 μM | SPR/SBA | Arac et al (2007) |
| NL1–36 | L399A/N400A/D402N/D571A/Q574A | ER | ND | ND | ND | SBA | This study |
| NL1–37 | L399A/N400A/D402N/F499A | PM | − | − | ND | SBA | This study |
| NL1–38 | T111A/Q112A | ER | ND | ND | ND | SBA | This study |
| NL1–39 | E92A/H93A | PM | + | + | ND | SBA | This study |
| NL1–40 | F458A/W463A | PM | + | + | ND | SBA | This study |
| NL1–41 | Q147A/D148A | PM | + | + | ND | SBA | This study |
| NL1–42 | R63A/K66A | ER | ND | ND | ND | SBA | This study |
| NL1–43 | S102A/S105A | PM | + | + | ND | SBA | This study |
| NL1–44 | F582A/K586A/R589A/E72Q/I73A | PM | + | + | ND | SBA | This study |
| NL1–45 | Y509A/H511A/Q513A/D421N/D423N/D424N | PM | + | + | ND | SBA | This study |
| NL1–46 | Q328A/Y332A/D440N/N441A/Y445A | PM | + | + | ND | SBA | This study |
| NL1–51 | F458A/M459A/W463A | PM | + | + | ND | SBA | This study |
| NL1–53 | F458A/M459A/W463A L629A/L633A | PM | + | + | ND | SBA | This study |
| NL1–54 | F458A/M459A/W463A/L399A/N400A/ D402N/E297A/K306A | ER | ND | ND | ND | ND | This study |
| NL1–ΔC | T776 followed by stop codon | PM | + | + | ND | SBA | This study |
| ER, endoplasmic reticulum; ITC, isothermal titration calorimetry; ND, not determined; NL1, neuroligin-1; PM, plasma membrane; SPR, surface plasmon resonance; SBA, surface binding assay (Figures 1, Supplementary Figure S4 and S5); all neuroligin-1 molecules used lacked inserts in splice sites A and B, and contained a cytoplasmic monomeric mVenus tag inserted between residues 776 and 777. | |||||||