Expression and characterization of catalytic domain of Plasmodium falciparum subtilisin-like protease 3

被引：11
作者：
Alam, Asrar [1 ]
Bhatnagar, Raj K. [2 ]
Chauhan, Virander S. [1 ]
机构：
[1] Int Ctr Genet Engn & Biotechnol, Malaria Res Grp, New Delhi 110067, India
[2] Int Ctr Genet Engn & Biotechnol, Insect Resistance Grp, New Delhi 110067, India
来源：
MOLECULAR AND BIOCHEMICAL PARASITOLOGY | 2012年 / 183卷 / 01期
基金：
美国国家卫生研究院;
关键词：
Plasmodium falciparum; Serine protease; Subtilisin-like protease; Erythrocytic schizogony; MALARIA PARASITE; IDENTIFICATION; CHEMOTHERAPY; INVASION; TARGETS; RELEASE; LIFE;
D O I：
10.1016/j.molbiopara.2011.12.009
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：
PfSUB3 is the third subtilisin-like protease annotated in Plasmodium genome database "PlasmoDB". The other two members, PfSUB1 and PfSUB2 have been implicated in merozoite egress and invasion in asexual blood stages. In this study, we recombinantly expressed a region of PfSUB3 spanning from Asn(334) to Glu(769) (PfSUB3c) which encompassed the predicted catalytic domain with all the active site residues and predicted mature region spanning from Thr(516) to Glu(769) (PfSUB3m) in E. coli. PfSUB3m showed PMSF-sensitive proteolytic activity in in vitro assays. Replacement of active site serine with alanine in PfSUB3m resulted in inactive protein. We found that PfSUB3c and PfSUB3m undergo truncation to produce a 25-kDa species which was sufficient for proteolytic activity. Quantitative real-time PCR, immnufluorescence assay and Western blot analyses revealed that PfSUB3 is expressed at late asexual blood stages. Serine protease activity of PfSUB3 and its expression in the late stages of erythrocytic schizogony are indicative of some possible role of the protease in merozoite egress and/or invasion processes. (C) 2012 Elsevier B.V. All rights reserved.
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收藏
页码：84 / 89
页数：6
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