Isolation and partial characterization of chromoprotein from the larval hemolymph of the Japanese oak silkworm (Antheraea yamamai).
A total of two different hemolymph proteins (designated P-I and P-II) of the Japanese oak silkworm, Antheraea yamamai, were purified from the hemolymph of the fifth instar larvae using four chromatographic steps: (a) hydrophobic interaction chromatography; (b) ion exchange chromatography; (c) gel-filtration; and (d) reverse-phase high performance liquid chromatography (HPLC). These two proteins were separated by TSKgel Phenyl-5PW RP column chromatography. P-I has an apparent molecular weight of 31,000 or 35,000, as determined by gel-filtration and SDS-PAGE, respectively. P-II shows a molecular weight of 22,000 or 25,000, by gel-filtration and SDS-PAGE, respectively. The molecular weight of P-I and P-II were determined to be 31,076 and 21,500 by MALDI-TOF MS, respectively. These results suggest that both P-I and P-II are monomers. The N-terminal sequence analysis suggests that P-I is closely related to the ommochrome-binding protein (OBP) from the hemolymph of Manduca sexta, with 40% identity in the first 30 residues, while P-II is similar to the biliproteins (BPs) from other lepidopteran insects (50% identity). Spectroscopic analysis shows that the blue chromophore of A. yamamai BP is not biliverdin IX, which is present in the biliproteins of most insects.
Saito
H
H
Department of Insect Genetics and Breeding, National Institute of Sericultural and Entomological Science, Ibaraki, Japan. saito13@ss.nises.affrc.go.jp
Yamada
H
H
Kato
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry; D006458:Hemolymph / Q000737:chemistry*; D019476:Insect Proteins / Q000737:chemistry*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009036:Moths / Q000196:embryology*; D016415:Sequence Alignment; D017421:Sequence Analysis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013053:Spectrophotometry
D002352:Carrier Proteins; D019476:Insect Proteins; C071652:OBP protein, Manduca sexta