Coenzyme A stimulates collagen production in cultured fibroblasts; possible mechanisms in enzymatic and gene expression.
Coenzyme A, a cofactor in enzymatic acetyl transfer reactions, stimulates collagen production in cultured fibroblasts. The mechanisms involved in this collagen stimulation were investigated. Enzymatic studies using radiolabeled procollagen as substrate revealed that coenzyme A enhanced prolyl hydroxylase activity. Prolyl hydroxylase is a key enzyme in collagen synthesis acting by hydroxylation of proline residues in procollagen peptide, which is necessary for stabilizing collagen. Northern blot analysis demonstrated that coenzyme A also enhanced mRNA levels of both the alpha subunit of prolyl hydroxylase and the alpha1 chain of type I collagen. The levels of protein production of prolyl hydroxylase and type I collagen were also increased in cultured fibroblasts by coenzyme A, which correlated well with observations from Northern blotting. On the other hand, coenzyme A did not stimulate the activity nor the gene expression of two other processing enzymes: lysyl hydroxylase, which provides the sites for glycosylation and crosslinking between collagen peptides, and lysyl oxidase, a fundamental enzyme in intermolecular crosslinking. These results indicate that coenzyme A stimulates collagen production by at least two separate mechanisms: by enhancing prolyl hydroxylase activity as well as stimulating gene expression of the alpha subunit of this enzyme, and by stimulating gene expression of alpha1 chain of type I collagen.
Mio
K
K
Life Science Research Center, Lion Corporation, Odawara, Kanagawa, Japan. kazumo@lion.co.jp
Yamashita
M
M
Odake
Y
Y
Tamai
H
H
Takada
K
K
D016475:3T3 Cells; D000818:Animals; D002478:Cells, Cultured; D003065:Coenzyme A / Q000494:pharmacology*; D003094:Collagen / Q000235:genetics; D024042:Collagen Type I / Q000096:biosynthesis*; D000090602:Collagen Type I, alpha 1 Chain; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007527:Isoenzymes / Q000235:genetics; D051379:Mice; D011393:Procollagen-Proline Dioxygenase / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats
D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D007527:Isoenzymes; D012333:RNA, Messenger; D003094:Collagen; D011393:Procollagen-Proline Dioxygenase; D003065:Coenzyme A