Journal ArticleDOI
Two-dimensional gel electrophoresis of membrane proteins
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TLDR
A high-resolution method for two-dimensional separation of membrane proteins is described, a modification of the one recently described by O'Farrell (1975); about 150 different proteins can be visualized in Escherichia coli or Salmonella typhimurim cell envelopes.Abstract:
A high-resolution method for two-dimensional separation of membrane proteins is described. It involves a nondiscriminating solubilization of a membrane preparation with sodium dodecyl sulfate, followed by electrophoresis in the first dimension according to charge (by isoelectric focusing). The electrophoresis in the second dimension is in the presence of sodium dodecyl sulfate, thus separating proteins on the basis of molecular weight. Electrophoresis in the first dimension is either on a thin slab gel, or on a small-diameter tube; electrophoresis in the second dimension is on a thin slab gel. Up to 100 mug of protein can be analyzed. The two-dimensional system is a modification of the one recently described by O'Farrell (1975). About 150 different proteins can be visualized in Escherichia coli or Salmonella typhimurim cell envelopes; examples of differences between mutant and wild-type strains are presented. The method is applicable also to membrane preparations from other sources: a two-dimensional separation of plasma membrane proteins from HeLa cells is presented.read more
Citations
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Journal ArticleDOI
Silver staining of proteins in polyacrylamide gels.
TL;DR: The silver-Staining procedure for detecting proteins in polyacrylamide gels has been modified and further simplified so that it is stable, controllable, and even more rapid than previous silver-staining methods.
Journal ArticleDOI
Identification of the 64K autoantigen in insulin-dependent diabetes as the GABA-synthesizing enzyme glutamic acid decarboxylase.
Steinunn Baekkeskov,Henk-Jan Aanstoot,Stephan Christgai,Annette Reetz,Michele Solimena,Marilia Cascalho,Franco Folli,Hanne Richter-Olesen,Pietro De Camilli +8 more
TL;DR: The pancreatic islet β-cell autoantigen of relative molecular mass 64,000 (64K), which is a major target of autoantibodies associated with the development of insulin-dependent diabetes mel-litus (IDDM), has been identified as glutamic acid decarboxylase, the biosynthesizing enzyme of the inhibitory neurotransmitter GABA.
Journal ArticleDOI
The nuclear envelope lamina is reversibly depolymerized during mitosis
Larry Gerace,Günter Blobel +1 more
TL;DR: It is demonstrated by immunofluorescence microscopy that the lamina is reversibly disassembled during cell division, coincident with the disassembly and reconstruction of the mitotic nuclear envelope architecture, and that all three lamins are monomeric at periods of mitotic lamina disassembly.
Journal ArticleDOI
Membrane proteins and proteomics: un amour impossible?
TL;DR: This review shows how two‐dimensional electrophoresis performs with membrane proteins from bacteria or animal or vegetable cells and tissues, the recent progress in this field, and it examines future prospects in this area.
Journal ArticleDOI
Solubilization of Plant Membrane Proteins for Analysis by Two-Dimensional Gel Electrophoresis
TL;DR: Gels of the best quality were obtained when membrane proteins were extracted with phenol and precipitated with ammonium acetate in methanol; background staining and streaking were diminished and proteins were clearly resolved.
References
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Journal Article
Protein Measurement with the Folin Phenol Reagent
TL;DR: Procedures are described for measuring protein in solution or after precipitation with acids or other agents, and for the determination of as little as 0.2 gamma of protein.
Journal ArticleDOI
The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis
Klaus Weber,Mary Osborn +1 more
TL;DR: The results show that the polyacrylamide gel electrophoresis method can be used with great confidence to determine the molecular weights of polypeptide chains for a wide variety of proteins.
Journal ArticleDOI
High resolution two-dimensional electrophoresis of proteins.
TL;DR: This technique provides a method for estimation of the number of proteins made by any biological system and can resolve proteins differing in a single charge and consequently can be used in the analysis of in vivo modifications resulting in a change in charge.
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Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane.
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Acetylornithinase of Escherichia coli: partial purification and some properties.
Henry J. Vogel,David M. Bonner +1 more
TL;DR: Compounds Used-N”l-Acetyl-n-ornithine was synthesized as previously described and L-Ornithine monohydrochloride was obtained from the Mann Research Laboratories.