| Title: | Reconstitution of SEC gene product-dependent intercompartmental protein transport |
| Author(s): | Baker D; Hicke L; Rexach M; Schleyer M; Schekman R; |
| Address: | "Department of Biochemistry, University of California, Berkeley 94720" |
| DOI: | 10.1016/0092-8674(88)90196-1 |
| ISSN/ISBN: | 0092-8674 (Print) 0092-8674 (Linking) |
| Abstract: | "Transport of alpha-factor precursor from the endoplasmic reticulum to the Golgi apparatus has been reconstituted in gently lysed yeast spheroplasts. Transport is measured through the coupled addition of outer-chain carbohydrate to [35S]methionine-labeled alpha-factor precursor translocated into the endoplasmic reticulum of broken spheroplasts. The reaction is absolutely dependent on ATP, stimulated 6-fold by cytosol, and occurs between physically separable sealed compartments. Transport is inhibited by the guanine nucleotide analog GTP gamma S. sec23 mutant cells have a temperature-sensitive defect in endoplasmic reticulum-to-Golgi transport in vivo. This defect has been reproduced in vitro using sec23 membranes and cytosol. Transport at 30 degrees C with sec23 membranes requires addition of cytosol containing the SEC23 (wild-type) gene product. This demonstrates that an in vitro inter-organelle transport reaction depends on a factor required for transport in vivo. Complementation of sec mutations in vitro provides a functional assay for the purification of individual intercompartmental transport factors" |
| Keywords: | Adenosine Triphosphate/pharmacology Biological Transport Cold Temperature Cytosol/metabolism Endoplasmic Reticulum/*metabolism Freezing Glycosylation Golgi Apparatus/*metabolism Guanosine 5'-O-(3-Thiotriphosphate) Guanosine Triphosphate/analogs & derivati; |
| Notes: | "MedlineBaker, D Hicke, L Rexach, M Schleyer, M Schekman, R eng Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. 1988/07/29 Cell. 1988 Jul 29; 54(3):335-44. doi: 10.1016/0092-8674(88)90196-1" |
