Chromaffin granule and PC12 cell chondroitin sulfate proteoglycans and their relation to chromogranin A.
Two major proteoglycans, which appear to be structurally closely related, were isolated from bovine chromaffin granule matrix proteins by ion-exchange chromatography. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis they have apparent average molecular sizes of 35-40 kDa (range of 23-75 kDa) and generate a 14-kDa core glycoprotein after chondroitinase treatment. Previous studies demonstrated that these two major chromaffin granule proteoglycans are very similar in terms of their peptide mapping patterns and carbohydrate composition (having a high proportion of tri- and tetraantennary N-glycosidic oligosaccharides, and O-glycosidic oligosaccharides consisting predominantly of disialyl derivatives of galactosyl(beta 1-3)N-acetylgalactosamine), and that they differed in these respects from the chromogranins. By using antisera to five synthetic peptide fragments of chromogranin A to stain immunoblots of purified chromaffin granule proteoglycans before and after chondroitinase treatment, we have now shown that these major proteoglycans are not immunochemically related to chromogranin A. However, it has recently been reported that some chromogranin A-immunoreactive material disappears after chondroitinase treatment, and our studies demonstrate that approximately 1-2% of the chromogranin A occurs in the form of a 110-kDa proteoglycan, which is converted to a 95-kDa core glycoprotein after chondroitinase treatment. Similar chromogranin A proteoglycans could be detected in rat PC12 pheochromocytoma cells, where they have a molecular size of 115-145 kDa and yield a 105-kDa core protein after chondroitinase treatment. Studies using antibodies to synthetic peptide fragments of chromogranin B (secretogranin I) did not provide any evidence that this related protein occurs in a proteoglycan form.
Gowda
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Department of Pharmacology, New York University Medical Center, New York 10016.
Hogue-Angeletti
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Margolis
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Margolis
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D000310:Adrenal Gland Neoplasms / Q000032:analysis*; D000818:Animals; D002417:Cattle; D011508:Chondroitin Sulfate Proteoglycans / Q000276:immunology / Q000302:isolation & purification*; D002837:Chromaffin Granules / Q000032:analysis*; D002838:Chromaffin System / Q000032:analysis*; D053379:Chromogranin A; D002864:Chromogranins / Q000032:analysis*; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D009419:Nerve Tissue Proteins / Q000032:analysis*; D010673:Pheochromocytoma / Q000032:analysis*; D011509:Proteoglycans / Q000302:isolation & purification*; D051381:Rats; D014407:Tumor Cells, Cultured / Q000032:analysis
D011508:Chondroitin Sulfate Proteoglycans; D053379:Chromogranin A; D002864:Chromogranins; D009419:Nerve Tissue Proteins; D011509:Proteoglycans; C506153:chromogranin A, rat