Mutations in the EBI2 major binding pocket
EBI2 mutations were assessed for their cell surface expression and ability to activate the CREB transcription factor in transiently transfected HEK293 cells. The table shows these values at 15 ng/8 × 104 cells and the relative constitutive activity (see Materials and Methods for calculation) at 15 and 25 ng/8 × 104 cells. CREB activity and cell surface expression are presented as percentage ± S.E.M., whereas the relative constitutive activity is presented as -fold ± S.E.M. In all cases, the results are given relative to the wt value. The number of experiments is given as n. Receptor residues are presented according to the Schwartz and Ballesteros/Weinstein (Ballest.) nomenclatures (see Introduction). Furthermore, the three most common amino acids at the particular position are given.
Residue | CREB Activity | Expression | Relative CA | n | |||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Mutation | Number | Schwartz | Ballest | Three Most Common AAs | 15 ng/well | 15 ng | 25 ng | ||||||||
| % | -fold | ||||||||||||||
| TM III | T(III:04)A | Thr107 | Thr III:04 | T3.28 | V, 19%; S, 10%; W, 10% | 97 ± 3.2 | 94 ± 1.6 | 1.0 ± 0.05 | 1.1 ± 0.07 | 3 | |||||
| T(III:04)E | 76 ± 12 | 53 ± 11 | 1.4 ± 0.08 | 1.2 ± 0.06 | 4 | ||||||||||
| F(III:08)A | Phe111 | Phe III:08 | F3.32 | D, 20%; F, 15%; L, 9% | 78 ± 10 | 108 ± 18 | 0.74 ± 0.05 | 0.82 ± 0.04 | 6 | ||||||
| F(III:08)Y | 73 ± 11 | 39 ± 6.4 | 1.9 ± 0.11 | 1.8 ± 0.14 | 6 | ||||||||||
| T(III:12)A | Thr115 | Thr III:12 | T3.36 | M, 20%; L, 15%; V, 11% | 116 ± 10 | 126 ± 31 | 0.93 ± 0.07 | 1.2 ± 0.09 | 3 | ||||||
| T(III:12)F | 142 ± 41 | 31 ± 91 | 0.46 ± 0.01 | 0.68 ± 0.09 | 4 | ||||||||||
| TM V | C(V:09)A | Cys201 | Cys V:09 | C5.43 | F, 23%; L, 13%; S, 10% | 45 ± 5.9 | 71 ± 7.5 | 0.63 ± 0.07 | 0.68 ± 0.07 | 4 | |||||
| G(V:12)W | Gly204 | Gly V:12 | C5.46 | G, 29%; L, 14%; A, 11% | 40 ± 2.7 | 66 ± 5.2 | 0.61 ± 0.03 | 0.64 ± 0.04 | 4 | ||||||
| Y(V:13)A | Tyr205 | Tyr V:13 | C5.47 | F, 70%; Y, 11%; L, 4% | 38 ± 4.7 | 54 ± 9.2 | 0.70 ± 0.08 | 0.64 ± 0.03 | 4 | ||||||
| Y(V:13)F | 86 ± 7.3 | 61 ± 14 | 1.4 ± 0.13 | 1.4 ± 0.10 | 4 | ||||||||||
| TM VI | F(VI:09)A | Phe253 | Phe VI:09 | F6.44 | F, 82%; Y, 6%; L, 3% | 75 ± 1.4 | 81 ± 8.9 | 0.91 ± 0.10 | 1.0 ± 0.07 | 5 | |||||
| C(VI:12)A | Cys256 | Cys VI:12 | C6.47 | C, 74%; S, 9%; F, 5% | 95 ± 1.8 | 41 ± 1.6 | 2.3 ± 0.21a | 2.4 ± 0.19a | 4 | ||||||
| F(VI:13)A | Phe257 | Phe VI:13 | F6.48 | W, 71%; F, 16%; Q, 2% | 39 ± 7.5 | 7.0 ± 1.1 | 5.7 ± 0.50b | 4.7 ± 0.27b | 5 | ||||||
| F(VI:13)C | 54 ± 8.3 | 17 ± 1.8 | 3.1 ± 0.27a | 4.8 ± 0.27b | 6 | ||||||||||
| F(VI:13)W | 67 ± 5.3 | 73 ± 11 | 0.9 ± 0.08 | 1.0 ± 0.13 | 4 | ||||||||||
| Y(VI:16)A | Tyr260 | Tyr VI:16 | Y6.51 | Y, 36%; F, 30%; L, 12% | 25 ± 3.6 | 57 ± 7.3 | 0.43 ± 0.04 | 0.53 ± 0.07 | 6 | ||||||
| Y(VI:16)F | 49 ± 4.7 | 96 ± 15 | 0.53 ± 0.03 | 0.59 ± 0.14 | 5 | ||||||||||
| M(VI:24)A | Met268 | Met VI:24 | M6.59 | A, 16%; T, 14%; L, 12% | 89 ± 4.7 | 109 ± 9.2 | 0.82 ± 0.07 | 0.84 ± 0.07 | 5 | ||||||
| TM VII | T(VII:05)A | Thr293 | Thr VII:05 | T7.38 | T, 24%; A, 15%; S, 13% | 99 ± 7.1 | 126 ± 8.0 | 0.87 ± 0.05 | 0.69 ± 0.13 | 6 | |||||
| V(VII:06)A | Val294 | Val VII:06 | V7.39 | L, 21%; V, 11%; T, 11% | 110 ± 11 | 88 ± 5.9 | 1.3 ± 0.07 | 1.4 ± 0.10 | 4 | ||||||
| M(VII:09)A | Met297 | Met VII:09 | M7.42 | A, 40%; G, 20%; S, 13% | 65 ± 2.3 | 93 ± 2.2 | 0.71 ± 0.02 | 0.78 ± 0.05 | 4 | ||||||
| M(VII:09)F | 85 ± 15 | 80 ± 2.9 | 1.1 ± 0.06 | 1.4 ± 0.15 | 4 | ||||||||||
|
| N(VII:12)A | Asn300 | Asn VII:12 | N7.45 | N, 67%; S, 12%; H, 9% | 103 ± 5.8 | 146 ± 20 | 0.73 ± 0.08 | 0.81 ± 0.11 | 3 | |||||