Isolation, characterization, and cDNA sequence of a carotenoid binding protein from the silk gland of Bombyx mori larvae

Hiroko Tabunoki, Hiromu Sugiyama, Yoshiro Tanaka, Hiroshi Fujii, Yutaka Banno, Zeina E. Jouni, Masahiko Kobayashi, Ryoichi Sato, Hideaki Maekawa, Kozo Tsuchida

研究成果: ジャーナルへの寄稿記事

66 引用 (Scopus)

抄録

A carotenoid binding protein (CBP) has been isolated from the silk glands of Bombyx mori larvae. The protein has an apparent molecular mass of 33 kDa and binds carotenoids in a 1:1 molar ratio. Lutein accounts for 90% of the bound carotenoids, whereas α-carotene and β-carotene are minor components. Immunological analysis demonstrated the presence of CBP only in the yellow-colored tissues of the silk gland, midgut, testis, and ovary. Several phenotypes of B. mori mutants linked to carotenoid transport have been utilized to characterize CBP. The Y (yellow hemolymph) gene controls uptake of carotenoids from the midgut lumen into the midgut epithelium, and larvae with the +Y gene lack this property. Immunoblotting analysis confirmed the presence of CBP in mutants with the dominant Y gene only. Immunohistochemistry verified the localization of CBP in the villi of the midgut epithelium, indicating that CBP might be involved in absorption of carotenoids. A cDNA clone for CBP encoding a protein of 297 amino acids has been isolated from the B. mori silk gland cDNA library. The deduced amino acid sequence revealed that CBP is a novel member of the steroidogenic acute regulatory (StAR) protein family with its unique structural feature of a StAR-related lipid transfer domain, known to aid in lipid transfer and recognition. Lutein-binding capacity of the recombinant CBP (rCBP) determined by incubating rCBP with lutein followed by immunoprecipitation using anti-CBP IgG conjugated to protein A-Sepharose, demonstrated the formation of a lutein-rCBP complex. Sequence analyses coupled with binding specificity suggest that CBP is a new member of the StAR protein family that binds carotenoids rather than cholesterol.

元の言語英語
ページ(範囲)32133-32140
ページ数8
ジャーナルJournal of Biological Chemistry
277
発行部数35
DOI
出版物ステータス出版済み - 8 30 2002

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Bombyx
Silk
Carotenoids
Larva
Carrier Proteins
Complementary DNA
Lutein
Genes
Epithelium
Lipids
Amino Acids
Dominant Genes
Hemolymph
Molecular mass

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

これを引用

Isolation, characterization, and cDNA sequence of a carotenoid binding protein from the silk gland of Bombyx mori larvae. / Tabunoki, Hiroko; Sugiyama, Hiromu; Tanaka, Yoshiro; Fujii, Hiroshi; Banno, Yutaka; Jouni, Zeina E.; Kobayashi, Masahiko; Sato, Ryoichi; Maekawa, Hideaki; Tsuchida, Kozo.

:: Journal of Biological Chemistry, 巻 277, 番号 35, 30.08.2002, p. 32133-32140.

研究成果: ジャーナルへの寄稿記事

Tabunoki, H, Sugiyama, H, Tanaka, Y, Fujii, H, Banno, Y, Jouni, ZE, Kobayashi, M, Sato, R, Maekawa, H & Tsuchida, K 2002, 'Isolation, characterization, and cDNA sequence of a carotenoid binding protein from the silk gland of Bombyx mori larvae', Journal of Biological Chemistry, 巻. 277, 番号 35, pp. 32133-32140. https://doi.org/10.1074/jbc.M204507200
Tabunoki, Hiroko ; Sugiyama, Hiromu ; Tanaka, Yoshiro ; Fujii, Hiroshi ; Banno, Yutaka ; Jouni, Zeina E. ; Kobayashi, Masahiko ; Sato, Ryoichi ; Maekawa, Hideaki ; Tsuchida, Kozo. / Isolation, characterization, and cDNA sequence of a carotenoid binding protein from the silk gland of Bombyx mori larvae. :: Journal of Biological Chemistry. 2002 ; 巻 277, 番号 35. pp. 32133-32140.
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title = "Isolation, characterization, and cDNA sequence of a carotenoid binding protein from the silk gland of Bombyx mori larvae",
abstract = "A carotenoid binding protein (CBP) has been isolated from the silk glands of Bombyx mori larvae. The protein has an apparent molecular mass of 33 kDa and binds carotenoids in a 1:1 molar ratio. Lutein accounts for 90{\%} of the bound carotenoids, whereas α-carotene and β-carotene are minor components. Immunological analysis demonstrated the presence of CBP only in the yellow-colored tissues of the silk gland, midgut, testis, and ovary. Several phenotypes of B. mori mutants linked to carotenoid transport have been utilized to characterize CBP. The Y (yellow hemolymph) gene controls uptake of carotenoids from the midgut lumen into the midgut epithelium, and larvae with the +Y gene lack this property. Immunoblotting analysis confirmed the presence of CBP in mutants with the dominant Y gene only. Immunohistochemistry verified the localization of CBP in the villi of the midgut epithelium, indicating that CBP might be involved in absorption of carotenoids. A cDNA clone for CBP encoding a protein of 297 amino acids has been isolated from the B. mori silk gland cDNA library. The deduced amino acid sequence revealed that CBP is a novel member of the steroidogenic acute regulatory (StAR) protein family with its unique structural feature of a StAR-related lipid transfer domain, known to aid in lipid transfer and recognition. Lutein-binding capacity of the recombinant CBP (rCBP) determined by incubating rCBP with lutein followed by immunoprecipitation using anti-CBP IgG conjugated to protein A-Sepharose, demonstrated the formation of a lutein-rCBP complex. Sequence analyses coupled with binding specificity suggest that CBP is a new member of the StAR protein family that binds carotenoids rather than cholesterol.",
author = "Hiroko Tabunoki and Hiromu Sugiyama and Yoshiro Tanaka and Hiroshi Fujii and Yutaka Banno and Jouni, {Zeina E.} and Masahiko Kobayashi and Ryoichi Sato and Hideaki Maekawa and Kozo Tsuchida",
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T1 - Isolation, characterization, and cDNA sequence of a carotenoid binding protein from the silk gland of Bombyx mori larvae

AU - Tabunoki, Hiroko

AU - Sugiyama, Hiromu

AU - Tanaka, Yoshiro

AU - Fujii, Hiroshi

AU - Banno, Yutaka

AU - Jouni, Zeina E.

AU - Kobayashi, Masahiko

AU - Sato, Ryoichi

AU - Maekawa, Hideaki

AU - Tsuchida, Kozo

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