Purification, cDNA cloning and expression of human NG,NG‐dimethylarginine dimethylaminohydrolase
Open Access
- 25 December 1998
- journal article
- research article
- Published by Wiley in European Journal of Biochemistry
- Vol. 258 (2) , 863-868
- https://doi.org/10.1046/j.1432-1327.1998.2580863.x
Abstract
CDNA encoding NG,NG‐dimethylarginine dimethylaminohydrolase from rat kidney had been cloned [Kimoto, M., Sasakawa, T., Tsuji, H., Miyatake, S., Oka, T., Nio, N. & Ogawa, T. (1997) Biochim. Biophys. Acta 1337, 6−10]. The enzyme hydrolyzes NG,NG‐dimethyl‐ L‐arginine and NG‐monomethyl‐ L‐arginine, which are known as endogenous inhibitors for the nitric oxide‐generating system. In the present study, human NG,NG‐dimethylarginine dimethylaminohydrolase has been purified to homogeneity from liver and characterized. The cDNA clone encoding human NG,NG‐dimethylarginine dimethylaminohydrolase was isolated from a human kidney λgt10 library using a probe prepared from a plasmid containing the entire coding region of rat NG,NG‐dimethylarginine dimethylaminohydrolase. Its open reading frame encoded a protein of 285 amino acids with a molecular mass of 31 121 Da. The deduced amino acid sequence exhibits 93 % identity with that of rat. The cDNA was expressed as a fusion protein in Escherichia coli and the recombinant protein exhibited enzyme activity which is the same as that of natural enzyme.Keywords
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