cDNA cloning and characterization of Type I procollagen alpha1 chain in the skate Raja kenojei.
A full-length cDNA of the Type I procollagen alpha1 [pro-alpha1(I)] chain (4388 bp), coding for 1463 amino acid residues in the total length, was determined by RACE PCR using a cDNA library constructed from 4-week embryo of the skate Raja kenojei. The helical region of the skate pro-alpha1(I) chain consisted of 1014 amino acid residues - the same as other fibrillar collagen alpha chains from higher vertebrates. Comparison on denaturation temperatures of Type I collagens from the skate, rainbow trout (Oncorhynchus mykiss) and rat (Rattus norvegicus) revealed that the number of Gly-Pro-Pro and Gly-Gly in the alpha1(I) chains could be directly related to the thermal stability of the helix. The expression property of the skate pro-alpha1(I) chain mRNA and phylogenetic analysis with other vertebrate pro-alpha1(I) chains suggested that skate pro-alpha1(I) chain could be a precursor form of the skate Type I collagen alpha1 chain. The present study is the first evidence for the primary structure of full-length pro-alpha1(I) chain in an elasmobranch.
Department of Marine Bioscience, Faculty of Biotechnology, Fukui Prefectural University, Obama, Fukui 917-0003, Japan.
Jae-Ho
JH
Hwang
Yoshihiro
Y
Yokoyama
Shoshi
S
Mizuta
Reiji
R
Yoshinaka
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D024042:Collagen Type I / Q000235:genetics* / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics*; D004622:Embryo, Mammalian / Q000737:chemistry / Q000378:metabolism; D004625:Embryo, Nonmammalian; D015723:Gene Library; D008969:Molecular Sequence Data; D010802:Phylogeny; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D012860:Skates, Fish / Q000235:genetics*; D013696:Temperature
D024042:Collagen Type I; D018076:DNA, Complementary