PURIFICATION AND IN VITRO ANALYSIS OF YEAST VACUOLES

Autor(en): Cabrera, Margarita
Ungermann, Christian 
Herausgeber: Klionsky, DJ
Stichwörter: ALPHA-SNAP; Biochemical Research Methods; Biochemistry & Molecular Biology; DOCKING; EARLY-STAGE; FUSION REQUIRES; GTPASE YPT7P; MEMBRANE-FUSION; PROTEIN; SACCHAROMYCES-CEREVISIAE; TRANS-SNARE COMPLEX; VERTEX RING DOMAIN
Erscheinungsdatum: 2008
Herausgeber: ELSEVIER ACADEMIC PRESS INC
Journal: AUTOPHAGY: LOWER EUKARYOTES AND NON-MAMMALIAN SYSTEMS, PT A
Methods in Enzymology
Volumen: 451
Startseite: 177
Seitenende: 196
Zusammenfassung: 
The purification of eukaryotic organelles is a prerequisite for the detailed analysis of protein sorting, localization and translocation, membrane fusion and vesicle budding. Yeast vacuoles receive cargo from the exocytic, endocytic, and autophagic pathways and hence represent an excellent model system for the study of organelle biogenesis and protein sorting. Yeast vacuoles undergo fission and fusion in vivo, events that can be monitored in vitro by an assay that employs purified vacuoles from two tester strains. Here, we describe the methodology of yeast vacuole purification, and provide protocols for the detailed analysis of the fusion reaction. We also include methods to analyze protein dynamics on yeast vacuoles and the controls required to ensure their reliability.
ISBN: 9780123745484
ISSN: 00766879
DOI: 10.1016/S0076-6879(08)03213-8

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