Purification and identification of a tributyltin-binding protein from serum of Japanese flounder, Paralichthys olivaceus

Yohei Shimasaki, Yuji Oshima, Yoshiko Yokota, Takeshi Kitano, Miki Nakao, Shun-Ichiro Kawabata, Nobuyoshi Imada, Tsuneo Honjo

研究成果: ジャーナルへの寄稿記事

30 引用 (Scopus)

抄録

Tributyltin (TBT) is an industrial chemical used as an antifoulant in marine environments. Previously, we reported that TBT accumulates in the serum or plasma of some fishes and is bound to a high molecular weight compound in the serum of the Japanese flounder Paralichthys olivaceus. In this study, we succeeded in purifying the TBT-binding protein (TBT-bp) from the serum of Japanese flounder by using gel filtration chromatography, anion exchange chromatography, and polyacrylamide gel electrophoresis, with a 2.6% yield and a 77-fold purification. The molecular mass of TBT-bp was approximately 46.5 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and its isoelectric point was approximately 3.0 on isoelectric focusing-polyacrylamide gel electrophoresis. The TBT-bp contained 42% N-glycan. The cDNA nucleotide sequence of TBT-bp was determined by reverse transcription-polymerase chain reaction of Japanese flounder liver, and we deduced a sequence of 191 amino acids of mature TBT-bp. No sequence identical to the TBT-bp amino acid sequence was found within the SWISS-PROT (http://www.nig. ac.jp/) protein database; however, a lipocalin-like sequence pattern was observed. We concluded that the TBT-bp was a novel protein that has not yet been reported, although some DNA sequences from expressed sequence tags (ESTs) of Japanese flounder liver had a high identity. A high expression level of TBT-bp gene was found in the liver, but the gene was slightly detectable in the kidney and brain.

元の言語英語
ページ(範囲)1229-1235
ページ数7
ジャーナルEnvironmental Toxicology and Chemistry
21
発行部数6
出版物ステータス出版済み - 1 1 2002

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Flounder
tributyltin
Purification
purification
serum
Carrier Proteins
protein
Serum
gel
Electrophoresis
Liver
Polyacrylamide Gel Electrophoresis
electrokinesis
Protein Databases
Chromatography
chromatography
Amino Acid Sequence
amino acid
Genes
Lipocalins

All Science Journal Classification (ASJC) codes

  • Environmental Chemistry
  • Health, Toxicology and Mutagenesis

これを引用

Purification and identification of a tributyltin-binding protein from serum of Japanese flounder, Paralichthys olivaceus. / Shimasaki, Yohei; Oshima, Yuji; Yokota, Yoshiko; Kitano, Takeshi; Nakao, Miki; Kawabata, Shun-Ichiro; Imada, Nobuyoshi; Honjo, Tsuneo.

:: Environmental Toxicology and Chemistry, 巻 21, 番号 6, 01.01.2002, p. 1229-1235.

研究成果: ジャーナルへの寄稿記事

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abstract = "Tributyltin (TBT) is an industrial chemical used as an antifoulant in marine environments. Previously, we reported that TBT accumulates in the serum or plasma of some fishes and is bound to a high molecular weight compound in the serum of the Japanese flounder Paralichthys olivaceus. In this study, we succeeded in purifying the TBT-binding protein (TBT-bp) from the serum of Japanese flounder by using gel filtration chromatography, anion exchange chromatography, and polyacrylamide gel electrophoresis, with a 2.6{\%} yield and a 77-fold purification. The molecular mass of TBT-bp was approximately 46.5 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and its isoelectric point was approximately 3.0 on isoelectric focusing-polyacrylamide gel electrophoresis. The TBT-bp contained 42{\%} N-glycan. The cDNA nucleotide sequence of TBT-bp was determined by reverse transcription-polymerase chain reaction of Japanese flounder liver, and we deduced a sequence of 191 amino acids of mature TBT-bp. No sequence identical to the TBT-bp amino acid sequence was found within the SWISS-PROT (http://www.nig. ac.jp/) protein database; however, a lipocalin-like sequence pattern was observed. We concluded that the TBT-bp was a novel protein that has not yet been reported, although some DNA sequences from expressed sequence tags (ESTs) of Japanese flounder liver had a high identity. A high expression level of TBT-bp gene was found in the liver, but the gene was slightly detectable in the kidney and brain.",
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AU - Shimasaki, Yohei

AU - Oshima, Yuji

AU - Yokota, Yoshiko

AU - Kitano, Takeshi

AU - Nakao, Miki

AU - Kawabata, Shun-Ichiro

AU - Imada, Nobuyoshi

AU - Honjo, Tsuneo

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