Prophenoloxidase-activating proteinase-3 (PAP-3) from Manduca sexta hemolymph:: a clip-domain serine proteinase regulated by serpin-1J and serine proteinase homologs

被引:194
作者
Jiang, HB [1 ]
Wang, Y
Yu, XQ
Zhu, YF
Kanost, M
机构
[1] Oklahoma State Univ, Dept Entomol & Plant Pathol, Stillwater, OK 74078 USA
[2] Univ Missouri, Sch Biol Sci, Div Cell Biol & Biophys, Kansas City, MO 64110 USA
[3] Kansas State Univ, Dept Biochem, Manhattan, KS 66506 USA
关键词
insect immunity; melanization; phenoloxidase; serine proteinase; clip domain; serpin; serine proteinase homolog; tobacco hornworm;
D O I
10.1016/S0965-1748(03)00123-1
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Phenoloxidase (PO) is a key enzyme implicated in several defense mechanisms in insects and crustaceans. It is converted from prophenoloxidase (proPO) through limited proteolysis by prophenoloxidase-activating proteinase (PAP). We previously isolated PAP-1 from integument and PAP-2 from hemolymph of the tobacco hornworm, Manduca sexta. Here, we report the purification, characterization. and regulation of PAP-3 from the hemolymph. Similar to M. sexta PAP-2, PAP-3 consists of two amino-terminal clip domains followed by a carboxyl-terminal catalytic domain, whereas PAP-1 contains only one clip domain at its amino-terminus. Purified PAP-3 cleaved proPO at Arg(51) and generated a low level of PO activity. However, the enzyme efficiently activated proPO when M. sexta serine proteinase homolog-1 and -2 were present. These proteinase-like proteins associate with immulectin-2, a pattern-recognition receptor for lipopolysaccharide. M. sexta PAP-3 was inhibited by recombinant serpin-1J, which formed an SDS-stable complex with the enzyme. PAP-3 mRNA was detected at a low level in the fat body or hemocytes of naive larvae, but was elevated in insects that had been challenged with bacteria. These data, along with our previous results on PAP-1 and PAP-2, indicate that proPO activation by PAPs is a tightly regulated process. Individual PAPs could play different roles during immune responses and developmental processes. (C) 2003 Elsevier Ltd. All rights reserved.
引用
收藏
页码:1049 / 1060
页数:12
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