EFFICIENT TRANSLOCATION OF POSITIVELY CHARGED RESIDUES OF M13 PROCOAT PROTEIN ACROSS THE MEMBRANE EXCLUDES ELECTROPHORESIS AS THE PRIMARY FORCE FOR MEMBRANE INSERTION

被引:58
作者
KUHN, A [1 ]
ZHU, HY [1 ]
DALBEY, RE [1 ]
机构
[1] OHIO STATE UNIV,DEPT CHEM,COLUMBUS,OH 43210
关键词
M13 coat protein; membrane insertion; sec dependence;
D O I
10.1002/j.1460-2075.1990.tb07413.x
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 coat protein of bacteriophage MN13 is inserted into the Escherichia coli plasma membrane as a precursor protein, termed precoat, with a typical leader peptide of 23 amino acid residues. Its membrane insertion requires the electrochemical potential but not the cellular components SecA and SecY. Since the electrochemical gradients result in the periplasmic side of the membrane being positively charged, the membrane potential could contribute to the transfer of the negatively charged central region of precoat across the membrane. Here we demonstrate that the central domain following the leader peptide can be translocated across the membrane even when the net charge of the region is changed from -3 to +3. This rules out an electrophoresis-like insertion mechanism for procoat. We also show that the sec independence of procoat insertion is linked to the presence of the second apolar domain. The deletion of most of the second apolar domain from a procoat fusion protein results in sec dependent membrane insertion of the hybrid protein. Moreover, like other proteins that require the sec genes, translocation of this sec dependent procoat protein is inhibited when positively charged residues are introduced after the leader peptide. Loop models involving one or two hydrophobic regions are presented that account for the differences in tolerance of positively charged residues.
引用
收藏
页码:2385 / 2389
页数:5
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