Purification and characterization of tributyltin-binding protein type 2 from plasma of Japanese flounder, Paralichthys olivaceus.
We used gel filtration chromatography, anion-exchange chromatography and polyacrylamide gel electrophoresis to purify tributyltin-binding protein type 2 (TBT-bp 2) from plasma of Japanese flounder (Paralichthys olivaceus) injected intraperitoneally with TBT (5.0 mg/kg body weight). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that the molecular mass of TBT-bp 2 was approximately 48 kDa, and isoelectric focusing-polyacrylamide gel electrophoresis indicated that the isoelectric point was approximately 3.0. TBT-bp 2 contained 40% N-glycan. The complete cDNA nucleotide sequence and the genome sequence of TBT-bp 2 were determined by means of rapid amplification of cDNA ends of liver tissue of Japanese flounder and a genome-walking technique, respectively. The 216 amino acid sequence of TBT-bp 2 showed 47% identity to the sequences of puffer fish (Takifugu pardalis) saxitoxin- and tetrodotoxin-binding protein but only 27% similarity to the sequence of TBT-bp 1. Analysis of the motif sequence of the amino acid sequence and the structure of the gene encoding TBT-bp 2 suggested that this protein belongs to the lipocalin superfamily.
Oba
Yumi
Y
Laboratory of Marine Environmental Science, Division of Marine Biological Chemistry, Department of Bioscience and Biotechnology, Kyushu University, Higashi-ku, Fukuoka, 812-8581, Japan.
Shimasaki
Yohei
Y
Oshima
Yuji
Y
Satone
Hina
H
Kitano
Takeshi
T
Nakao
Miki
M
Kawabata
Shun-ichiro
S
Honjo
Tsuneo
T
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002850:Chromatography, Gel; D018076:DNA, Complementary / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D029941:Fish Proteins / Q000097:blood* / Q000737:chemistry* / Q000302:isolation & purification; D005432:Flounder; D016678:Genome; D007564:Japan; D054834:Lipocalins / Q000097:blood* / Q000737:chemistry* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D014220:Trialkyltin Compounds / Q000008:administration & dosage / Q000378:metabolism*
D018076:DNA, Complementary; D029941:Fish Proteins; D054834:Lipocalins; D014220:Trialkyltin Compounds; C011559:tributyltin