Identification, expression, and tissue distribution of the three rat lysyl hydroxylase isoforms.
Lysyl hydroxylases (LH) (procollagen-lysine 2-oxoglutarate 5-dioxygenase; PLOD) catalyse the hydroxylation of lysine residues during the post-translational modification of collagenous proteins. In this paper, we describe the first identification and cloning of LH isoforms 2 and 3 from the rat, including both LH2 splice variants (LH2a and LH2b). The rat LHs are expressed in almost all tissue and cell types examined, indicating a probable lack of tissue specificity for LH function. All LH isoforms were stably transfected into CHO-K1 cells and this represents the first example of recombinant LH production in a eukaryotic cell line. Expression and production of all LH isoforms led to an increase in total collagen synthesis. LH1 and LH2a expression and production led to an increase in total pyridinium cross-link production. Evidence that LH2a possesses telopeptide lysyl hydroxylase activity, previously thought to be a novel enzyme, is presented.
Mercer
Derry K
DK
Rowett Research Institute, Greenburn Road, Bucksburn, Aberdeen AB21 9SB, UK. D.Mercer@rowett.ac.uk
Nicol
Phyllis F
PF
Kimbembe
Cisca
C
Robins
Simon P
SP
D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003094:Collagen / Q000096:biosynthesis / Q000378:metabolism; D024042:Collagen Type I; D006224:Cricetinae; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D010455:Peptides / Q000378:metabolism; D008240:Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase / Q000235:genetics* / Q000378:metabolism*; D012326:RNA Splicing; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D020539:Sequence Analysis, Protein; D014018:Tissue Distribution; D014158:Transcription, Genetic
D024042:Collagen Type I; D007527:Isoenzymes; D010455:Peptides; D011994:Recombinant Proteins; C052929:collagen type I trimeric cross-linked peptide; D003094:Collagen; D008240:Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase