Production and characterization of monoclonal antibodies to porcine brain pyridoxal kinase.
Six monoclonal antibodies that recognize porcine brain pyridoxal kinase have been selected and designated as PK67, PK86, PK91, PK144, PK252 and PK275. A total of six monoclonal antibodies recognizing different epitopes of the enzyme were obtained, of which four inhibited the enzyme activity. When total proteins of porcine brain homogenate separated by SDS-PAGE were subjected to monoclonal antibodies, a single reactive protein band of molecular weight 39 kDa which comigrated with purified porcine pyridoxal kinase was detected. Using the anti-pyridoxal kinase antibodies as probes, the cross reactivities of brain pyridoxal kinase from human and other mammalian tissues and from avian sources were also investigated. Among human and all animal tissues tested, immunoreactive bands on Western blots appeared to have the same molecular mass of 39 kDa. These results indicate that mammalian brains contain only one major type of immunologically similar pyridoxal kinase, although some properties of the enzymes reported previously differed from one another.
Choi
S Y
SY
Department of Genetic Engineering, Hallym University, Chunchon, Korea. sychoi@sun.hallym.ac.kr
Kwok
F
F
Bahn
J H
JH
Jeon
S G
SG
Ahn
Y K
YK
Yoon
B H
BH
Lee
B R
BR
Choi
K S
KS
Gao
G Z
GZ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000918:Antibody Specificity; D001921:Brain / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000276:immunology; D005260:Female; D005909:Glioblastoma / Q000201:enzymology; D006801:Humans; D015151:Immunoblotting; D051379:Mice; D008807:Mice, Inbred BALB C; D009447:Neuroblastoma / Q000201:enzymology; D011731:Pyridoxal Kinase / Q000032:analysis / Q000276:immunology*; D013552:Swine; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000939:Epitopes; D011731:Pyridoxal Kinase