The number of amino acid residues in hydrophilic loops connecting transmembrane domains of the GABA transporter GAT‐1 is critical for its function

FEBS Letters

Su, Hailing¹  Pantanowitz, Shifrah¹  Bendahan, Annie¹  Kanner, Baruch I.¹ 

[1] Department of Biochemistry, Hadassah Medical School, The Hebrew University, PO Box 12272, Jerusalem 91120, Israel

关键词: GABA transport;    Site-directed mutagenesis;    Immunoprecipitation;    Reconstitution;    Rat brain;    GABA;    γ-aminobutyric acid;    loop n−(n + 1);    the hydrophilic loop connecting putative transmembrane α-helices n and n + 1;    ΔX-Y;    deletion of amino acids from X to Y;   

DOI  :  10.1016/0014-5793(94)01255-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Transporter proteins consist of multiple transmembrane domains connected by hydrophillic loops. As the importance of these loops in transport processes is poorly understood, we have studied this question using the cDNA coding for GAT-1, a Na⁺/Cl⁻-coupled γ-aminobutyric acid transporter from rat brain. Deletions of randomly picked non-conserved single amino acids in the loops connecting helices 7 and 8 or 8 and 9 result in inactive transport upon expression in HeLa cells. However, transporters where these amino acids are replaced with glycine retain significant activity. The expression levels of the inactive mutant transporters was similar to that of the wild-type, but one of these, ΔVal-348, appears to be defectively targetted to the plasma membrane. Our data are compatible with the idea that a minimal length of the loops is required, presumably to enable the transmembrane domains to interact optimally with each other.

【 授权许可】

Unknown