Serine protease SP7 cleaves prophenoloxidase and is regulated by two serpins in Ostrinia furnacalis melanization.

Abstract

Melanization is an innate immune response in insects to defend against the invading pathogens and parasites. During melanization, prophenoloxidase (PPO) requires proteolytic activation by its upstream prophenoloxidase-activating protease (PAP). We here cloned a full-length cDNA for a serine protease, named as SP7, from Ostrinia furnacalis. The open reading frame of SP7 encodes 421-amino acid residue protein with a 19-residue signal peptide. qRT-PCR analysis showed that SP7 mRNA levels were significantly upregulated upon exposure to microbial infection. Recombinant SP7 zymogen was activated by serine protease SP2. The active SP7 could cleave O. furnacalis PPOs including PPO2, PPO1b and PPO3. Additionally, active SP7 could form covalent complexes with serine protease inhibitor serpin-3 and serpin-4. The activity of SP7 in cleaving a colorimetric substrate IEARpNA or O. furnacalis PPOs was efficiently blocked by either serpin-3 or serpin-4. Our work thus revealed that SP7 and SP2 partially constituted a PPO activation cascade in which SP7 was activated by SP2 and then likely worked as a PAP. SP7 was effectively regulated by serpin-3 and serpin-4. The results would allow further advances in the understanding of melanization mechanisms in O. furnacalis.

• Authors: Shasha Zhang, Ting Feng, Jiayue Ji, Lei Wang, Chunju An
• Journal: Insect biochemistry and molecular biology (Insect Biochem Mol Biol) Vol. 141 Pg. 103699 (02 2022) ISSN: 1879-0240 [Electronic] England
• PMID: 34920078 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2021. Published by Elsevier Ltd.

Chemical References

• Enzyme Precursors
• Insect Proteins
• Serpins
• pro-phenoloxidase
• Catechol Oxidase
• Serine Proteases

Topics

• Animals
• Catechol Oxidase (metabolism)
• Enzyme Precursors (metabolism)
• Insect Proteins (genetics, metabolism)
• Larva (enzymology, genetics, growth & development, metabolism)
• Moths (enzymology, genetics, growth & development, metabolism)
• Serine Proteases (genetics, metabolism)
• Serpins (genetics, metabolism)