Functional and physical interactions between partial molecules of STE6, a yeast ATP-binding cassette protein

被引:24
作者
Berkower, C [1 ]
Taglicht, D [1 ]
Michaelis, S [1 ]
机构
[1] JOHNS HOPKINS SCH MED, DEPT ANAT & CELL BIOL, BALTIMORE, MD 21205 USA
关键词
D O I
10.1074/jbc.271.38.22983
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The Saccharomyces cerevisiae a-factor transporter, STE6, is a member of the ATP binding cassette (ABC) transporter superfamily, ABC proteins consist of four modular units that comprise two membrane spanning domains (MSDs) and two nucleotide-binding domains (NBDs). Like many ABC proteins, STE6 contains these four domains in a single polypeptide; certain other ABC proteins are encoded as pairs of "half-molecules" or are further subdivided. Our previous studies demonstrated that STE6 can be expressed as two half-molecules that are functional when co-expressed, Here we dissect the interactions between modules of STE6 in greater detail, We show by co-immunoprecipitation that STE6 half-molecules interact physically, supporting the view that they co-assemble in vivo to form a functional transporter, We also demonstrate a physical interaction between a STE6 half-molecule and full-length STE6; such complexes appear to be functional, based on the striking finding that the defective activity of full-length STE6 mutated in one of its NBDs can be corrected by co-expression of the corresponding "wild-type" half-molecule. We also show that a quarter-molecule consisting solely of the N-terminal MSD of STE6 can interact physically and functionally with a C-terminal three-quarter molecule of STE6, indicating that information directing the assembly of STE6 from partial molecules is contained, at least in part, within its membrane spans.
引用
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页码:22983 / 22989
页数:7
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