DOI: 10.1083/jcb.120.1.95. Reconstitution of protein translocation from solubilized yeast membranes reveals topologically distinct roles for BiP and cytosolic Hsc70.

Reconstitution of protein translocation from solubilized yeast membranes reveals topologically distinct roles for BiP and cytosolic Hsc70.

10.1083/jcb.120.1.95

Crossref journal-article

Rockefeller University Press

The Journal of cell biology (291)

Abstract

We reconstituted prepro-alpha-factor translocation and signal peptide processing using a yeast microsomal detergent soluble fraction formed into vesicles with soybean phospholipids. Reconstituted translocation required ATP, and was deficient when sec63 and kar2 (BiP) mutant cells were used as a source of membranes. Normal translocation was observed with vesicles reconstituted from a mixture of pure wild-type yeast BiP and a soluble fraction of kar2 mutant membranes. Two other heat-shock cognate (hsc) 70 homologs, yeast cytosolic hsc70 (Ssalp) and E. coli dnaK protein did not replace BiP. Conversely, BiP was not active under conditions where translocation into native ER vesicles required cytosolic hsc70. We conclude that cytosolic hsc70 and BiP serve noninterchangeable roles in polypeptide translocation, possibly because distinct, asymmetrically oriented membrane proteins are required to recruit each protein to opposing surfaces of the ER membrane.

Bibliography

Brodsky, J. L., Hamamoto, S., Feldheim, D., & Schekman, R. (1993). Reconstitution of protein translocation from solubilized yeast membranes reveals topologically distinct roles for BiP and cytosolic Hsc70. The Journal of Cell Biology, 120(1), 95â102.

Authors 4

J L Brodsky (first)
S Hamamoto (additional)
D Feldheim (additional)
R Schekman (additional)

References 0 Referenced 136

None

Dates

Type	When
Created	21 years, 3 months ago (May 14, 2004, 8:22 p.m.)
Deposited	2 years, 1 month ago (July 21, 2023, 10:51 p.m.)
Indexed	11 months, 3 weeks ago (Sept. 6, 2024, 9:16 p.m.)
Issued	32 years, 8 months ago (Jan. 1, 1993)
Published	32 years, 8 months ago (Jan. 1, 1993)
Published Online	32 years, 8 months ago (Jan. 1, 1993)
Published Print	32 years, 8 months ago (Jan. 1, 1993)

Funders 0

None

BibTeX

@article{Brodsky_1993, title={Reconstitution of protein translocation from solubilized yeast membranes reveals topologically distinct roles for BiP and cytosolic Hsc70.}, volume={120}, ISSN={1540-8140}, url={http://dx.doi.org/10.1083/jcb.120.1.95}, DOI={10.1083/jcb.120.1.95}, number={1}, journal={The Journal of cell biology}, publisher={Rockefeller University Press}, author={Brodsky, J L and Hamamoto, S and Feldheim, D and Schekman, R}, year={1993}, month=jan, pages={95–102} }

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