RRC ID |
451
|
著者 |
Tabunoki H, Sugiyama H, Tanaka Y, Fujii H, Banno Y, Jouni ZE, Kobayashi M, Sato R, Maekawa H, Tsuchida K.
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タイトル |
Isolation, characterization, and cDNA sequence of a carotenoid binding protein from the silk gland of Bombyx mori larvae.
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ジャーナル |
J Biol Chem
|
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 alpha-carotene and beta-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.
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巻・号 |
277(35)
|
ページ |
32133-40
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公開日 |
2002-8-30
|
DOI |
10.1074/jbc.M204507200
|
PII |
S0021-9258(20)70074-8
|
PMID |
12052833
|
MeSH |
Amino Acid Sequence
Animals
Base Sequence
Bombyx / genetics*
Bombyx / physiology
Carrier Proteins / analysis
Carrier Proteins / genetics*
Carrier Proteins / metabolism
DNA, Complementary / chemistry
DNA, Complementary / genetics
Exocrine Glands / metabolism*
Insect Proteins / analysis
Insect Proteins / genetics*
Insect Proteins / metabolism
Larva
Molecular Sequence Data
Molecular Weight
Mutagenesis
Sequence Alignment
Sequence Homology, Amino Acid
|
IF |
4.238
|
引用数 |
59
|
WOS 分野
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
リソース情報 |
カイコ |
Y strains ? |