Purification and characterization of tributyltin-binding protein of tiger puffer, Takifugu rubripes.
We successfully purified Trub.TBT-bpα, a tributyltin (TBT) binding protein (bp) of the tiger puffer, Takifugu rubripes. Tiger puffer was injected intraperitoneally with TBT (1.0mg/kg body weight) and Trub.TBT-bpα was purified from serum by ammonium sulfate fractionation, gel filtration chromatography and polyacrylamide gel electrophoresis. Gel electrophoresis revealed that the Trub.TBT-bpα has a molecular mass of approximately 48.5kDa and contains at least 40% N-glycan. The deduced 212 amino acid sequence of the protein showed the highest identity (41%, 212 amino acid overlap and E-value: 9e-42) with TBT-binding protein type 1 (TBT-bp1) of Paralichthys olivaceus (Japanese flounder). Analysis of the gene structure of Trub.TBT-bpα suggests that this protein belongs to the lipocalin superfamily, which may be important in the accumulation and elimination of TBT. Phylogenetic analysis suggests that functionalization of TBT-bps has occurred during evolution, and that the functions of this group of proteins might be important for fish survival.
Oba
Yumi
Y
Laboratory of Marine Environmental Science, Faculty of Agriculture, Kyushu University, Hakozaki 6-10-1, Higashi-ku, Fukuoka, 812-8581, Japan. ooba-y@agr.kyushu-u.ac.jp
Yamauchi
Akira
A
Hashiguchi
Yasuyuki
Y
Satone
Hina
H
Miki
Shizuho
S
Nassef
Mohamed
M
Shimasaki
Yohei
Y
Kitano
Takeshi
T
Nakao
Miki
M
Kawabata
Shun-ichiro
S
Honjo
Tsuneo
T
Oshima
Yuji
Y
D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000097:blood / Q000737:chemistry* / Q000302:isolation & purification; D052244:Endocrine Disruptors / Q000378:metabolism*; D029941:Fish Proteins / Q000097:blood / Q000737:chemistry* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010802:Phylogeny; D023741:Takifugu / Q000097:blood / Q000378:metabolism*; D014220:Trialkyltin Compounds / Q000378:metabolism*
D002352:Carrier Proteins; D052244:Endocrine Disruptors; D029941:Fish Proteins; D014220:Trialkyltin Compounds; C011559:tributyltin