Occurrence of two distinct types of tissue inhibitor of metalloproteinases-2 in teleost fish.
We have cloned for the first time two cDNAs encoding distinct types of tissue inhibitor of metalloproteinases-2 (TIMP-2) from teleost fish, Japanese flounder, and designated these types as jfTIMP-2a and jfTIMP-2b. The open reading frames of the jfTIMP-2a and jfTIMP-2b cDNAs are composed of 663 and 657 nucleotides and 221 and 219 amino acids, respectively. Both jfTIMP-2s contain 12 cysteine residues, which might form six disulfide bonds as in other animals' TIMP-2s. The predicted full-length amino acid sequence of jfTIMP-2a has lower identity to jfTIMP-2b (63%) than to those of human (74%) and chicken (73%) TIMP-2s, but higher than to those of other human TIMPs (TIMP-1: 39%, TIMP-3: 43%, TIMP-4: 45%), indicating that jfTIMP-2a is a common TIMP-2, while jfTIMP-2b is unique to Japanese flounder. However, the C-terminal region including the last three disulfide bonds of jfTIMP-2b has higher amino acid identity to those of other animal TIMP-2s than to that of jfTIMP-2a. Reverse-transcribed polymerase chain reaction (RT-PCR) analysis showed the mRNAs of jfTIMP-2a and jfTIMP-2b to be ubiquitously expressed in all tissues examined, but with different expression patterns. These findings suggest that the two distinct jfTIMP-2s might perform different functions in teleost tissues.
Kubota
Satoshi
S
Department of Aquaculture, Kochi University, Nankoku 783-8502, Japan. kubota@cc.kochi-u.ac.jp
Kinoshita
Masato
M
Uji
Susumu
S
Yokoyama
Yoshihiro
Y
Yamamoto
Eiichi
E
Hirono
Ikuo
I
Aoki
Takashi
T
Sakaguchi
Morihiko
M
Morioka
Katsuji
K
Itoh
Yoshiaki
Y
Toyohara
Haruhiko
H
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D005399:Fishes / Q000378:metabolism*; D005432:Flounder / Q000378:metabolism; D015870:Gene Expression; D008969:Molecular Sequence Data; D010802:Phylogeny; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000235:genetics*
D019716:Tissue Inhibitor of Metalloproteinase-2