Modulation of protease activity to enhance the recovery of recombinant nucleocapsid protein of Nipah virus

被引:6
|
作者
Chong, Fui Chin [1 ,2 ]
Tan, Wen Siang [3 ,4 ]
Biak, Dayang Radiah Awang [1 ]
Ling, Tau Chuan [4 ,5 ]
Tey, Beng Ti [1 ,4 ]
机构
[1] Univ Putra Malaysia, Dept Chem & Environm Engn, Fac Engn, Serdang 43400, Selangor, Malaysia
[2] Univ Malaysia Pahang, Dept Chem & Nat Resources Engn, Fac Engn, Kuantan 25000, Pahang, Malaysia
[3] Univ Putra Malaysia, Dept Microbiol, Fac Biotechnol & Biomol Sci, Serdang 43400, Selangor, Malaysia
[4] Univ Putra Malaysia, Inst Biosci, Serdang 43400, Selangor, Malaysia
[5] Univ Putra Malaysia, Dept Proc & Food Engn, Fac Engn, Serdang 43400, Selangor, Malaysia
关键词
Endogenous protease; Protease inhibitor; Nucleocapsid protein; Nipah virus; Escherichia coli; ESCHERICHIA-COLI; CLEAVAGE; INHIBITION; CYSTEINE; HENDRA; SITE;
D O I
10.1016/j.procbio.2009.08.012
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
The nucleocapsid (N) protein of Nipah virus (NiV) expressed in Escherichia coli (E. coli) is antigenic and immunogenic. A method to enhance the recovery of recombinant N protein of NiV produced in E. coli is described. A bioinformatics tool, PeptideCutter was used to identify potential protease and cleavage sites from the amino acid sequences deduced from the published DNA sequence of the N protein of NiV. The size of degraded protein was estimated by using the Western blot and PeptideCutter analyse. The identified proteases were serine proteases, hence, a range of serine protease inhibitors were tested to improve the recovery of the N protein. The relative amount of N protein of NiV was 2-fold higher with the addition of PMSF, compared to the control sample (without any protease inhibitor supplementation). (C) 2009 Elsevier Ltd. All rights reserved.
引用
收藏
页码:133 / 137
页数:5
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