Research article summary (published 29 Sep 2009):

Expression, purification, and characterization of pro-phenoloxidase-activating serine protease from Spodoptera litura.

Full Abstract

One of the important trigger molecules for innate immunity is a serine protease that activates zymogen phenol oxidase (PPO). Central to wound healing response is the activation of phenol oxidase zymogen. Molecular characterization of phenol oxidase has been recently reported by us. Here, we report isolation, cloning, expression, and purification of prophenol oxidase activating enzyme 1 (slppae1) from polyphagous pest, Spodoptera litura. SLPPAE1 is induced within 6 h of physical injury. The structural features of the mature polypeptide are reminiscent of other lepidopteran PPAE in having a signal peptide, propeptide, and catalytically active polypeptide. The cDNA has been expressed in Sf21 cells using baculovirus expression vector. Fractionation of expressing Sf21 cells revealed its expression in the membranes. The recombinant protein was solubilized from membranes and purified by Ni-NTA affinity chromatography. The purified enzyme is catalytically active on chromogenic substrate, activates recombinantly expressed prophenol oxidase (PPO) of S. litura, and is sensitive to inhibition by aprotenin. N-terminal sequencing of processed phenol oxidase revealed 11 kDa propeptide instead of in-silico predicted 6 kDa polypeptide.

Author information

Author/s: Arora, Naresh (N); Hoque, M E (ME); Rajagopal, R (R); Sachdev, Bindiya (B); Bhatnagar, Raj K (RK);

Affiliation: Insect Resistance Group, International Centre for Genetic Engineering and Biotechnology (ICGEB), New Delhi, India.

Journal and publication information

Publication Type: Journal Article

Journal: Archives of insect biochemistry and physiology (Arch Insect Biochem Physiol), published in United States. (Language: eng)

Reference: 2009-Oct; vol 72 (issue 2) : pp 61-73

Dates: Created 2009/09/09; Completed 2009/10/28;

PMID: 19557749, status: MEDLINE (last retrieval date: 10/28/2009, IMS Date: )

Sourced from the National Library of Medicine. Abstract text and other information may be subject to copyright.

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MeSH headings (categories)

This article was linked to the MESH Headings shown below.

Amino Acid Sequence Animals Cloning, Molecular Enzyme Activation Gene Expression Regulation, Enzymologic - genetics Molecular Sequence Data

Molecular Sequence Data Monophenol Monooxygenase - metabolism Phylogeny Sequence Alignment Serine Endopeptidases - biosynthesis; genetics; isolation & purification Spodoptera - enzymology

Associated Chemicals: Monophenol Monooxygenase (EC 1.14.18.1) ; Serine Endopeptidases (EC 3.4.21.-) ; prephenoloxidase-activating enzyme (EC 3.4.21.-)

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