Vesicle-Mediated Protein Sorting

Pryer, Nancy; Wuestehube, Linda J.; Schekman, Randy

doi:10.1146/annurev.bi.61.070192.002351

Cited by 454 publications

(271 citation statements)

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“…The elaboration and imaginative use of cell-free systems for the study ofintracellular trafficking events has galvanized this field of cell biology, leading to rapid advances in our understanding of the molecular basis of these events [22][23][24][25][26][27][28]. The genetic approach to the study of trafficking and sorting has allowed for the identification of a myriad of proteins implicated in these events in yeast [29], and it has already proved possible in a few instances to identify the mammalian counterpart protein.…”

Section: Protein Secretory Pathways: a General Outline Leader And Leamentioning

confidence: 99%

Sorting and processing of secretory proteins

Halban

Irminger

1994

Biochemical Journal

307

225

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Section: Protein Secretory Pathways: a General Outline Leader And Leamentioning

confidence: 99%

Sorting and processing of secretory proteins

Halban

Irminger

1994

Biochemical Journal

307

225

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“…The former phe-DISCUSSION We have described a new method for isolating mammalian cells with changes in secretory activity. The usefulness of the genetic approach in understanding secretion is already established from the work in Saccharomyces cerevisiae where the large collection of secretory mutants is yielding important information on the components that are involved in this pathway (Pryer et al, 1992). Although it is possible that differences in the basic mechanisms which control vesicle formation and consumption in yeast and mammalian cells might be minor, we expect that in mammals additional mechanisms may exist which couple secretion to signal transduction and perhaps provide different cell types with a more specialized repertoire of control mechanisms.…”

Section: Isolation Of Cho Cells With Defects In Secretionmentioning

confidence: 99%

A fluorescent lipid analogue can be used to monitor secretory activity and for isolation of mammalian secretion mutants.

Ktistakis

Kao

Wang

et al. 1995

MBoC

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The use of reporter proteins to study the regulation of secretion has often been complicated by posttranslational processing events that influence the secretion of certain proteins, but are not part of the cellular mechanisms that specifically regulate secretion. This has been a particular limitation for the isolation of mammalian secretion mutants, which has typically been a slow process. To provide a reporter of secretory activity independent of protein processing events, cells were labeled with the fluorescent lipid analogue C5-DMB-ceramide (ceramide coupled to the fluorophore boron dipyrromethene difluoride) and its secretion was followed by fluorescence microscopy and fluorescenceactivated cell sorting. Brefeldin A, which severely inhibits secretion in Chinese hamster ovary cells, blocked secretion of C5-DMB-ceramide. At high temperature, export of C5-DMB-ceramide was inhibited in HRP-1 cells, which have a conditional defect in secretion. Using C5-DMB-ceramide as a reporter of secretory activity, several different pulse-chase protocols were designed that selected mutant Chinese hamster ovary cells that were resistant to the drug brefeldin A and others that were defective in the transport of glycoproteins to the cell surface. Mutant cells of either type were identified in a mutagenized population at a frequency of 10-6. Thus, the fluorescent lipid C5-DMBceramide can be used as a specific marker of secretory activity, providing an efficient, general approach for isolating mammalian cells with defects in the secretory pathway.

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“…NSF and SNAPs were originally detected as factors required for transport through the Golgi in in vitro assays [6,7,21] and the yeast homologues of these proteins, secl8 and secl7, are essential for secretion in vivo [22]. In Golgi transport assays and in the formation of a Golgi membrane derived 20S complex, a-and ]3-SNAP appear to be functionally redundant whereas y-SNAP binds to different membrane sites and has a distinct role in stabilisation of the 20S complex [23,24].…”

Section: Introductionmentioning

confidence: 99%

Similar effects of α‐ and β‐SNAP on Ca ²⁺‐regulated exocytosis

et al. 1996

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We have examined this possibility by comparing the activities of recombinant a-and ~SNAPs. Both of these proteins were able to similarly bind NSF and activate its ATPase activity but to a lesser extent than y-SNAP. When introduced into digitoninpermeabilised chromaffin cells, both a-and J~-SNAP stimulated Ca2+-regulated exocytosis in a MgATP-dependent manner. The dose-response relationships for these proteins were essentially the same and addition of both proteins did not lead to any further increase in exocytosis above that due to each protein alone. We conclude that a-and/]-SNAPs are interchangeable isoforms with similar functions in regulated exocytosis.

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Vesicle-Mediated Protein Sorting

Cited by 454 publications

References 0 publications

Sorting and processing of secretory proteins

Sorting and processing of secretory proteins

A fluorescent lipid analogue can be used to monitor secretory activity and for isolation of mammalian secretion mutants.

Similar effects of α‐ and β‐SNAP on Ca ²⁺‐regulated exocytosis

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Vesicle-Mediated Protein Sorting

Cited by 454 publications

References 0 publications

Sorting and processing of secretory proteins

Sorting and processing of secretory proteins

A fluorescent lipid analogue can be used to monitor secretory activity and for isolation of mammalian secretion mutants.

Similar effects of α‐ and β‐SNAP on Ca 2+‐regulated exocytosis

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Similar effects of α‐ and β‐SNAP on Ca ²⁺‐regulated exocytosis