Human liver catalase: cloning, expression and characterization of monoclonal antibodies.
We isolated a cDNA encoding liver catalase from a human liver cDNA library. The cDNA had a high degree of sequence similarity to the corresponding enzyme from other sources. It was expressed in E. coli using the pET15b vector. The protein produced was enzymatically active after purification, and its kinetic parameters closely resembled those of other mammalian catalases. Monoclonal antibodies were generated against the purified catalase; six antibodies recognizing different epitopes were obtained, one of which inhibited the enzyme. The cross reactions of the antibodies with brain catalases from human and other mammalian tissues were investigated, and all the immunoreactive bands obtained on Western blots had molecular masses of about 58 kDa. Similarly fractionated extracts of several mammalian cell lines all gave a single band of molecular mass 58 kDa. These results indicate that mammalian livers and human cell lines contain only one major type of immunologically reactive catalase, even though some of catalases have been previously reported to differ in certain properties.
Jin
Li Hua
LH
Division of Life Sciences, Hallym University, Chunchon 200-702, Korea.
Kim
Dae Won
DW
Eum
Won Sik
WS
Yoon
Chang Sik
CS
Jang
Sang Ho
SH
Choi
Hee Soon
HS
Choi
Soo Hyun
SH
Kim
Young Hoon
YH
Kim
So Young
SY
Jung
Mi Ryoung
MR
Kang
Tae-Cheon
TC
Won
Moo Ho
MH
Lee
Hyeon Yong
HY
Kang
Jung Hoon
JH
Kwon
Oh-Shin
OS
Cho
Sung-Woo
SW
Lee
Kil Soo
KS
Park
Jinseu
J
Choi
Soo Young
SY
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics*; D002374:Catalase / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D002460:Cell Line; D003001:Cloning, Molecular; D003429:Cross Reactions; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D006801:Humans; D008099:Liver / Q000201:enzymology*; D051379:Mice; D008969:Molecular Sequence Data; D051381:Rats; D016415:Sequence Alignment
D000911:Antibodies, Monoclonal; D002374:Catalase