Characterization of acyl-CoA-binding protein (ACBP) in the pheromone gland of the silkworm, Bombyx mori.
Various fatty acyl-CoAs are involved as intermediates or precursors of sex pheromone components in the biosynthetic pathway of the pheromones in many lepidopteran insects. We have purified a 10-kDa protein from the cytosolic fraction of Bombyx mori pheromone glands by using affinity chromatography with a palmitoyl-CoA-agarose column and reversed-phase HPLC. Amino acid sequence analysis of the fragment peptides obtained from the purified protein, and a homology search, revealed that this protein was a member of acyl-CoA-binding proteins (ACBPs). MALDI-TOF mass spectral analysis of the purified protein and cloning of the gene from a pheromone gland cDNA library confirmed B. mori ACBP to be a 90 amino acid protein with 78.9% identity to that of Manduca sexta ACBP. The secondary structure of the recombinant B. mori ACBP was determined by NMR spectroscopy. Northern blot analysis demonstrated that B. mori ACBP was predominantly expressed in the pheromone gland and the corresponding transcript was expressed from the day before adult eclosion. Present results suggest that ACBP plays a significant role in the production of sex pheromones regulated by the neurohormone, pheromone biosynthesis activating neuropeptide (PBAN).
Matsumoto
S
S
The Institute of Physical and Chemical Research (RIKEN), Hirosawa 2-1, Wako-shi, Saitama 351-0198, Japan. smatsu@postman.riken.go.jp
Yoshiga
T
T
Yokoyama
N
N
Iwanaga
M
M
Koshiba
S
S
Kigawa
T
T
Hirota
H
H
Yokoyama
S
S
Okano
K
K
Mita
K
K
Shimada
T
T
Tatsuki
S
S
D000214:Acyl Coenzyme A; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern / Q000379:methods; D012831:Bombyx / Q000737:chemistry*; D002352:Carrier Proteins / Q000032:analysis* / Q000235:genetics; D002417:Cattle; D018076:DNA, Complementary; D026261:Diazepam Binding Inhibitor; D006801:Humans; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid; D012724:Sex Attractants
D000214:Acyl Coenzyme A; D002352:Carrier Proteins; D018076:DNA, Complementary; D026261:Diazepam Binding Inhibitor; D012724:Sex Attractants