Use of two-stage incubations to define sequential intermediates in endoplasmic reticulum to Golgi transport.
Identification of the temporal requirement for components through the use of two-stage incubations is valuable in dissecting the overall transport reaction into steps relevant to vesicle fission and those related to vesicle fusion. In the context of semiintact mammalian cells in which a functional vesicle intermediate has not been detected, components playing a role in targeting are presently difficult to identify. However, the two-stage incubations are particularly powerful when either the donor or acceptor compartments can be manipulated independently, as is the case for intra-Golgi transport using enriched Golgi fractions or in the case of ER-to-Golgi transport in perforated yeast, in which a vesicle intermediate can be physically isolated.
Davidson
H W
HW
Department of Cell and Molecular Biology, Scripps Research Institute, La Jolla, California 92037.
Balch
W E
WE
D001692:Biological Transport; D002451:Cell Compartmentation; D002463:Cell Membrane Permeability; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism*; D008562:Membrane Glycoproteins; D011499:Protein Processing, Post-Translational; D013997:Time Factors; D014759:Viral Envelope Proteins / Q000378:metabolism*; D015003:Yeasts / Q000378:metabolism*
C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins