title: Dopamine-beta-hydroxylase: structural comparisons of membrane-bound versus soluble forms from adrenal medulla and pheochromocytoma.
abstract: Dopamine-beta-hydroxylase (DBH) in membrane-bound (mDBH) and water-soluble (sDBH) forms was isolated from chromaffin granules of bovine adrenal medullae and a human pheochromocytoma tumor. sDBH was purified by concanavalin A-agarose column chromatography followed by DEAE-Sepharose column chromatography. The final bovine preparation had a specific activity of 16.27 IU/mg; the human preparation had a specific activity of 9.16 IU/mg. mDBH was isolated in enzymatically inactive form by preparative polyacrylamide gel electrophoresis. The proteins were subjected to amino acid analysis, as well as digestion with trypsin, followed by separation of the resulting peptides by two-dimensional TLC/electrophoresis. No intraspecies differences between sDBH and mDBH were found from comparisons of amino acid composition or peptide maps. Thus the basis of the difference between sDBH and mDBH cannot easily be explained by differences in primary structure, within the resolution of these techniques.
mesh_terms: D000310:Adrenal Gland Neoplasms / Q000201:enzymology*; D000313:Adrenal Medulla / Q000201:enzymology*; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000201:enzymology; D002837:Chromaffin Granules / Q000201:enzymology; D002845:Chromatography; D003600:Cytosol / Q000201:enzymology; D004299:Dopamine beta-Hydroxylase / Q000032:analysis / Q000378:metabolism*; D006801:Humans; D008970:Molecular Weight; D010673:Pheochromocytoma / Q000201:enzymology*; D014357:Trypsin
chemical_list: D000596:Amino Acids; D004299:Dopamine beta-Hydroxylase; D014357:Trypsin