Purification of aromatic L-amino acid decarboxylase from bovine brain with a monoclonal antibody.
Aromatic L-amino acid decarboxylase was purified from bovine brain for the first time by affinity chromatography using a monoclonal antibody to the enzyme, and it was compared with the decarboxylase purified from bovine adrenal medulla by the same procedure. The monoclonal antibody was produced from a hybridoma established for the enzyme highly purified from bovine adrenal medulla. The Mr values of brain and adrenal-medulla enzyme were both estimated to be approx. 100,000 by gel-permeation chromatography. SDS/polyacrylamide-gel electrophoresis revealed a single band with an apparent Mr of 50,000. Western immunoblot analysis showed that the antibody recognized each enzyme. With regard to substrate specificity, pH-dependence and effect of pyridoxal 5'-phosphate as a cofactor, both enzymes were similar.
Nishigaki
I
I
Department of Biochemistry, Nagoya University School of Medicine, Japan.
Ichinose
H
H
Tamai
K
K
Nagatsu
T
T
D000313:Adrenal Medulla / Q000201:enzymology; D000818:Animals; D000911:Antibodies, Monoclonal; D001142:Aromatic-L-Amino-Acid Decarboxylases / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D001921:Brain / Q000201:enzymology*; D002417:Cattle; D011232:Chemical Precipitation; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D007122:Immunoelectrophoresis; D007668:Kidney / Q000201:enzymology
D000911:Antibodies, Monoclonal; D001142:Aromatic-L-Amino-Acid Decarboxylases