Expression and purification of polyhistidine-tagged rotavirus NSP4 proteins in insect cells

被引：15

作者：

Rodríguez-Díaz, J

López-Andújar, P

García-Díaz, A

Cuenca, J

Montava, R

Buesa, J ^{[1
]}

机构：

[1] Univ Valencia, Sch Med, Dept Microbiol, E-46010 Valencia, Spain

[2] Univ Valencia, Hosp Clin Univ, E-46010 Valencia, Spain

来源：

PROTEIN EXPRESSION AND PURIFICATION | 2003年 / 31卷 / 02期

关键词：

rotavirus; NSP4; baculovirus; insect cells; NONSTRUCTURAL GLYCOPROTEIN; GENETIC GROUPS; SEQUENCE; ENTEROTOXIN; RESPONSES; PARTICLE; MODEL; SERUM; VP6;

D O I：

10.1016/S1046-5928(03)00166-9

中图分类号：

Q5 [生物化学];

学科分类号：

070307 [化学生物学];

摘要：

The rotavirus nonstructural NSP4 protein, a transmembrane endoplasmic reticulum-specific glycoprotein, has been described as the first viral enterotoxin. Purified NSP4 or a peptide corresponding to NSP4 residues 114-135 induces diarrhea in young mice. NSP4 has a membrane-destabilizing activity and causes an increase in intracellular calcium levels and chloride secretion by a calcium-dependent signalling pathway in eucaryotic cells. In this study, four recombinant baculoviruses were generated expressing the rotavirus NSP4 glycoprotein from the human strains Wa and Ito, the porcine strain OSU, and the simian strain SA11, which belong to two different NSP4 genotypes, A and B. The recombinant glycoproteins, expressed as polyhistidine-tagged molecules, were analyzed by Western blotting and immunoprecipitation. Newborn mice responded with diarrhea after inoculation with each of the recombinant NSP4 proteins. (C) 2003 Elsevier Science (USA). All rights reserved.

引用

页码：207 / 212

页数：6

共 24 条

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