A monoclonal antibody to dopamine beta-monooxygenase: detection of biosynthetic intermediates.
A monoclonal antibody to dopamine beta-monooxygenase (DBH) has been produced by an in vitro immunization technique. This antibody has been found to react with an epitope common to both soluble DBH (SDBH) and membrane-bound DBH (MDBH). A single CNBr fragment contains this antigenic site. Examination of electrophoretograms of chromaffin granule membrane and lysate by the immunoblot procedure revealed additional complexity. A 58-KDa polypeptide in low abundance was stained which could not be detected with the polyclonal antiserum. Chemical deglycosylation of SDBH produces two new polypeptides of 67 and 58 KDa. The entirety of this data suggests that the 58-KDa band is an unglycosylated form of DBH, proteolytically cleaved after biosynthesis.
J
J
Nolan
R
R
Fonseca
R H
RH
Angeletti
D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000915:Antibody Affinity; D000918:Antibody Specificity; D002837:Chromaffin Granules / Q000201:enzymology; D002999:Clone Cells; D003101:Collodion; D003488:Cyanogen Bromide; D004299:Dopamine beta-Hydroxylase / Q000096:biosynthesis / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D007120:Immunochemistry; D066298:In Vitro Techniques; D009638:Norepinephrine / Q000096:biosynthesis
D000911:Antibodies, Monoclonal; D000939:Epitopes; D003101:Collodion; D004299:Dopamine beta-Hydroxylase; D003488:Cyanogen Bromide; D009638:Norepinephrine