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

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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.

Original languageEnglish
Pages (from-to)32133-32140
Number of pages8
JournalJournal of Biological Chemistry
Volume277
Issue number35
DOIs
Publication statusPublished - Aug 30 2002

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Tabunoki, H., Sugiyama, H., Tanaka, Y., Fujii, H., Banno, Y., Jouni, Z. E., 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, 277(35), 32133-32140. https://doi.org/10.1074/jbc.M204507200