Purification and characterization of tributyltin-binding protein type 2 from plasma of Japanese flounder, Paralichthys olivaceus

Yumi Oba, Yohei Shimasaki, Yuji Oshima, Hina Satone, Takeshi Kitano, Miki Nakao, Shun-Ichiro Kawabata, Tsuneo Honjo

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

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 (5.0mg/kg body weight). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that the molecular mass of TBT-bp 2 was approximately 48kDa, 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.

Original languageEnglish
Pages (from-to)229-238
Number of pages10
JournalJournal of biochemistry
Volume142
Issue number2
DOIs
Publication statusPublished - Aug 1 2007

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

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