TY - JOUR
T1 - A monoclonal antibody to dopamine β-monooxygenase
T2 - Detection of biosynthetic intermediates
AU - Nolan, Judith
AU - Fonseca, Randi
AU - Angeletti, Ruth Hogue
N1 - Funding Information:
‘This work was supported by a grant from the National Institutes of Health (NS-13201). J.N. is supported by Training Grant NS-07064. a To whom correspondence should be addressed. 3 Abbreviations used: DBH, dopamine p-monooxygenase; SDBH, soluble DBH; MDBH, membrane-bound DBH; ELISA, enzyme-linked immunosorbent assay; PBS, phosphate-buffered saline (15 mM NaCI, 10 mM potassium phosphate, pH 7.2); DE52, diethylaminoethyl cellulose; BSA, bovine serum albumin; TBS, Tris-buffered saline (10 mM Tris, pH 7.4); SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; Hepes, 4-(Z-hydroxyethyl)-l-piperazineethanesulfonic acid; HAT, hypoxathine-aminopterin-thymidine.
PY - 1985/7
Y1 - 1985/7
N2 - A monoclonal antibody to dopamine β-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.
AB - A monoclonal antibody to dopamine β-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.
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U2 - 10.1016/0003-9861(85)90031-1
DO - 10.1016/0003-9861(85)90031-1
M3 - Article
C2 - 2409921
AN - SCOPUS:0021888597
SN - 0003-9861
VL - 240
SP - 257
EP - 264
JO - Archives of Biochemistry and Biophysics
JF - Archives of Biochemistry and Biophysics
IS - 1
ER -