Human liver catalase: cloning, expression and characterization of monoclonal antibodies.
Jin LH,
Kim DW,
Eum WS,
Yoon CS,
Jang SH,
Choi HS,
Choi SH,
Kim YH,
Kim SY,
Jung MR,
Kang TC,
Won MH,
Lee HY,
Kang JH,
Kwon OS,
Cho SW,
Lee KS,
Park J,
Choi SY.
Division of Life Sciences, Hallym University, Chunchon 200-702, Korea.
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.
PMID: 12872997 [PubMed - indexed for MEDLINE]