Rat lysyl hydroxylase: molecular cloning, mRNA distribution and expression in a baculovirus system.
A cDNA library from rat lung was screened with a chicken lysyl hydroxylase cDNA, and several overlapping rat lysyl hydroxylase cDNAs were isolated. The complete cDNA was 91 and 77% identical, respectively, to the human and chicken lysyl hydroxylase cDNAs at the protein level. By Northern blot, the rat lysyl hydroxylase cDNA recognized a single 3.2 kb mRNA that was present in a wide variety of rat tissues. In order to further confirm the identity of this cDNA, the cDNA was expressed in insect cells via a baculovirus vector. These cells produced an 85 kDa protein with lysyl hydroxylase activity. The recombinant lysyl hydroxylase had a specific activity and Km values for its substrates that were similar to those of the enzyme isolated from chick embryos. The fact that this single lysyl hydroxylase cDNA encodes a protein sufficient for lysyl hydroxylase activity is consistent with previous biochemical findings that lysyl hydroxylase only requires a single type of subunit for its activity.
Armstrong
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LC
Department of Internal Medicine, University of California, Davis, USA.
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D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000378:metabolism; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000096:biosynthesis* / Q000737:chemistry; D015870:Gene Expression; D005822:Genetic Vectors; D007313:Insecta; D008969:Molecular Sequence Data; D008240:Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics; D016415:Sequence Alignment
D018076:DNA, Complementary; D012333:RNA, Messenger; D011994:Recombinant Proteins; D008240:Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase