A group of integral membrane proteins of the rat liver Golgi contains a conserved protein of 100 kDa.
Rat liver Golgi membranes were washed with KCl and urea, and a polyclonal antiserum that stained the Golgi complex by immunofluorescence microscopy was raised. A group of proteins of apparent molecular mass 500 kDa, 200 kDa and 100 kDa were identified by immunoblotting with the antiserum, and were enriched in the Golgi membrane fraction. These proteins were also localised to the Golgi by immunofluorescence microscopy with affinity-purified antibodies. They are integral membrane proteins, and protease digestion experiments suggested that they are not exposed on the cytoplasmic face of the Golgi membrane. Immunofluorescence microscopy showed that staining of the Golgi complex by antibodies to the 100 kDa Golgi protein can be demonstrated among a wide range of mammalian species. This conservation may point to an important structural or functional role for the molecule. When the 100 kDa protein was reduced with dithiothreitol it was no longer recognised by the anti-Golgi antiserum. During phase separation in Triton X-114 the 100 kDa protein partitioned into the aqueous phase, rather than into the detergent phase, suggesting that it has a large luminal domain of hydrophilic amino acids.
Pryde
J G
JG
Department of Biochemistry, University of Edinburgh, Scotland, UK.
D000818:Animals; D015415:Biomarkers; D015153:Blotting, Western; D002458:Cell Fractionation; D016764:Cell Polarity; D002478:Cells, Cultured; D003429:Cross Reactions; D004229:Dithiothreitol / Q000494:pharmacology; D000939:Epitopes; D005260:Female; D005455:Fluorescent Antibody Technique; D005700:Galactosyltransferases / Q000032:analysis; D006056:Golgi Apparatus / Q000737:chemistry* / Q000187:drug effects / Q000276:immunology / Q000648:ultrastructure; D006801:Humans; D008099:Liver / Q000737:chemistry* / Q000166:cytology / Q000276:immunology / Q000648:ultrastructure; D008565:Membrane Proteins / Q000187:drug effects / Q000276:immunology / Q000302:isolation & purification*; D051381:Rats; D011916:Rats, Inbred F344; D013045:Species Specificity
D015415:Biomarkers; D000939:Epitopes; D008565:Membrane Proteins; D005700:Galactosyltransferases; D004229:Dithiothreitol