Molecular cloning of cDNA for pro-phenol-oxidase-activating factor i, a serine protease is induced by lipopolysaccharide or 1,3-β-glucan in coleopteran insect, Holotrichia diomphalia larvae

So Young Lee, Mi Young Cho, Ji Hoon Hyun, Kwang Moon Lee, Ko Ichi Homma, Shunji Natori, Shun-Ichiro Kawabata, Sadaaki Iwanaga, Bok Luel Lee

Research output: Contribution to journalArticlepeer-review

106 Citations (Scopus)

Abstract

Previously, we identified two pro-phenol oxidase-activating factors, named PPAF-I and PPAF-II, directly involved in the activation of the purified pro-phenol oxidase (pro-PO) from the hemolymph of the coleopteran, Holotrichia diomphalia larvae [Lee, S. Y., Kwon, T. H., Hyun, J. H., Choi, J. S., Kawabata, S. I., Iwanga, S, and Lee, B. L. (1998) Eur. J. Biochem. 254, 90-97]. Here, we report molecular cloning of cDNA for PPAF-I. Based on the sequence of the cloned cDNA, the PPAF-I gene appears to encode a member of serine protease zymogen consisting of 365 amino acid residues with a molecular mass of 40193 Da. The 109 amino acid residues preceding the amino- terminus Ile residue of the mature protein seem to constitute a prepro- sequence. The mature protein is a serine protease composed of 256 amino acids with a calculated molecular mass of 28009 Da. The overall structure is highly similar to that of Drosophila easter serine protease (42.9% identity), an essential serine protease zymogen for pattern formation in normal embryonic development. The locations of disulfide linkages in the pro-segment of PPAF- I were similar to those of Tachypleus proclotting enzyme and the mammalian neutrophil-derived defensin. Furthermore, [3H]diisopropylphosphate (iPr2P)- labeled PPAF-I was specifically produced from the crude preparation of PPAF- I zymogen by incubation with lipopolysaccharide or 1,3-β-glucan, whereas [3H]iPr2P-labeled PPAF-I was not produced under the same conditions in the absence of these microbial polysaccharides. These results indicate that the pro-PO-activation system in H. diomphalia larvae may proceed with the activation of PPAF-I zymogen by microbial polysaccharides.

Original languageEnglish
Pages (from-to)615-621
Number of pages7
JournalEuropean Journal of Biochemistry
Volume257
Issue number3
DOIs
Publication statusPublished - Nov 1 1998

All Science Journal Classification (ASJC) codes

  • Biochemistry

Fingerprint Dive into the research topics of 'Molecular cloning of cDNA for pro-phenol-oxidase-activating factor i, a serine protease is induced by lipopolysaccharide or 1,3-β-glucan in coleopteran insect, Holotrichia diomphalia larvae'. Together they form a unique fingerprint.

Cite this