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Methods Enzymol

Title:		Use of sec mutants to define intermediates in protein transport from endoplasmic reticulum
Author(s):		Rexach MF; Schekman RW;
Address:		"Division of Biochemistry and Molecular Biology, Howard Hughes Medical Research Institute, University of California, Berkeley 94720"
Journal Title:		Methods Enzymol
Year:		1992
Volume:		219
Issue:
Page Number:		267 - 286
DOI:		10.1016/0076-6879(92)19028-5
ISSN/ISBN:		0076-6879 (Print) 0076-6879 (Linking)
Abstract:		"In this chapter we have discussed the methodology used to identify and characterize three intermediates in protein transport from the ER that represent stages of transport vesicle budding, targeting, and fusion. The intermediates are obtained using a variety of transport inhibitors: low-temperature incubations, addition of chemicals, or inactivation of Sec protein function using temperature-sensitive mutants or specific antibodies. In all cases, the transport block imposed by the inhibitor is reversible, permitting assessment of the requirements for transport from each intermediate stage. Based on the differential requirements for Sec protein function, as well as the distinct kinetics and efficiency of transport from each intermediate, we demonstrate that each transport intermediate is functionally distinct"
Keywords:		Biological Transport/drug effects/genetics Calcium/pharmacology Cell Compartmentation Cell Membrane Permeability Cold Temperature Endoplasmic Reticulum/*metabolism Golgi Apparatus/*metabolism Kinetics Mating Factor Membrane Fusion Mutation Peptides/*metab;
Notes:		"MedlineRexach, M F Schekman, R W eng Comparative Study 1992/01/01 Methods Enzymol. 1992; 219:267-86. doi: 10.1016/0076-6879(92)19028-5"