Organized into seven parts encompassing 20 chapters, this volume begins with an overview of protein topology in the ER and the use of cross-linking methods to probe the molecular environment of translocating polypeptide chains. It then discusses the reconstitution of secretory protein translocation from detergent-solubilized rough microsomes; the use of anti-idiotype antibodies to characterize protein-protein interactions; the use of perforated cells to elucidate intracellular membrane transport; delivery of macromolecules into cells expressing a viral membrane fusion protein; and digitonin permeabilization procedures for studying endosome acidification and function. The reader is also introduced to reconstitution of intracellular vesicle fusion in a cell-free system after receptor-mediated endocytosis; immunoisolation using magnetic solid supports; endosome and lysosome purification by free-flow electrophoresis; remodeling of glycoprotein oligosaccharides after endocytosis; and replica plating of animal cells.
This book will interest students, researchers, geneticists, biochemists, and cell biologists.
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