ABCMdb

ABCB4 p.Gly581Ala

Predicted by SNAP2:	A: D (63%), C: D (63%), D: D (71%), E: D (75%), F: D (71%), H: D (71%), I: D (71%), K: D (75%), L: D (71%), M: D (53%), N: D (53%), P: D (80%), Q: D (63%), R: D (75%), S: D (63%), T: D (59%), V: D (66%), W: D (80%), Y: D (75%),
Predicted by PROVEAN:	A: D, C: D, D: D, E: D, F: D, H: D, I: D, K: D, L: D, M: D, N: D, P: D, Q: D, R: D, S: D, T: D, V: D, W: D, Y: D,

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Publications
[hide] Functional asymmetry of the two nucleotide binding... Mol Gen Genet. 2001 Feb;264(6):883-93. Proff C, Kolling R
Functional asymmetry of the two nucleotide binding domains in the ABC transporter Ste6.
Mol Gen Genet. 2001 Feb;264(6):883-93., [PMID:11254136]

Abstract [show]
The yeast a-factor transporter Ste6 is a member of the ABC transporter family and is closely related to human MDR1. We constructed a set of 26 Ste6 mutants using a random mutagenesis approach. Cell fractionation experiments demonstrated that most of the mutants, with the notable exception of those with alterations in TM1, are transported to the plasma membrane, the presumptive site of action of Ste6. Trafficking, therefore, does not seem to be affected in most of the mutants. To identify regions in Ste6 that interact with the ABC transporter "signature motif" (LSGGQ) we screened for intragenic revertants of the LSGGQ mutant M68 (S507N). Suppressor mutations were identified in TM12 and upstream of TM6. Surprisingly, these mutations also suppressed the Walker A mutation G397D, which should be defective in ATP-binding and hydrolysis at NBD1. Photoaffinity labeling experiments with 8-azido-[alpha-32P]ATP showed that ATP binding at NBD2 is reduced by the suppressor mutation in TM12. The experiments further suggest that the two NBDs of Ste6 are not equivalent and affect each other's ability to bind and hydrolyze ATP.

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Sentences [show]
No. Sentence Comment
96 Mutationa Motif b Mating eciencyc Nucleotide exchange Amino acid exchange 36 G580A, G581A, G588A, G591A G38N TM1 0.06 271 G580A, G581A G38N TM1 0.01 52 G581A G38D TM1 <10±6 263 G581A G38D TM1 0.06 319 G581A G38D TM1 <10±6 115 C675T, C676T, C679T Q70STOP, L71F TM1 0.15 86 G1427T G320V TM1 6.00 234 G1427A G320D TM1 5.50 285 G1642A, G1643A G392K WA <10±6 57 G1643A G392E WA <10±6 63 G1643A G392E WA <10±6 17 G1658A G397D WA <10±6 35 G1658A G397D WA <10±6 77 G1658A G397D WA <10±6 194 G1658A G397D WA 0.01 296 G1657A, G1658A G397N WA 1.7 24 G1783A E439 K Loop2 0.01 68 G1988A S507 N LSGGQ 0.50 197 G1988A S507 N LSGGQ 0.85 206 G1988A, G1990A, G1993A S507N, G508S, G509R LSGGQ <10±6 258 G2017C A517P WB 0.01 326 C2042T P525L WB 7.0 (25°C), 0.6 (30°C), <10±6 (37°C) 8 G2059A, G2065A E531K, V533I WB 0.75 110 C2363T, C2365T S632F, Q633STOP WB 1.8 349 G2966A, G3290A R833K, R941H WB 1.9 107 G3728A G1087D WA <10±6 44 G3743A G1092E WA <10±6 a Nucleotide positions are indicated according to GenBank Accession No. M26376 b TM1, transmembrane domain 1; WA/WB, Walker A and B motifs (Walker et al. 1982); Loop2 was de®ned by Hyde et al. (1990); LSGGQ, signature motif c Average mating activity based on at least two independent quantitative mating experiments. Login to comment