In vitro assembly into virus-like particles is an intrinsic quality of Pichia pastoris derived HCV core protein

Acosta-Rivero, N; Rodriguez, A; Musacchio, A; Falcón, V; Suarez, VM; Martinez, G; Guerra, I; Paz-Lago, D; Morera, Y; de la Rosa, MC; Morales-Grillo, J; Dueñas-Carrera, S

Abstract

Different variants of hepatitis C virus core protein (HCcAg) have proved to self-assemble in vitro into virus-like particles (VLPs). However, difficulties in obtaining purified mature HCcAg have limited these studies. In this study, a high degree of monomeric HCcAg purification was accomplished using chromatographic procedures under denaturing conditions. Size exclusion chromatography and sucrose density gradient centrifugation of renatured HCcAg (in the absence of structured RNA) under reducing conditions suggested that it assembled into empty capsids. The electron microscopy analysis of renatured HCcAg showed the presence of spherical VLPs with irregular shapes and an average diameter of 35 nm. Data indicated that HCcAg monomers assembled in vitro into VLPs in the absence of structured RNA, suggesting that recombinant HCcAg used in this work contains all the information necessary for the assembly process. However, they also suggest that some cellular factors might be required for the proper in vitro assembly of capsids. (C) 2004 Elsevier Inc. All rights reserved.

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Título según WOS: ID WOS:000225173400010 Not found in local WOS DB
Título de la Revista: BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Volumen: 325
Número: 1
Editorial: ACADEMIC PRESS INC ELSEVIER SCIENCE
Fecha de publicación: 2004
Página de inicio: 68
Página final: 74
DOI:

10.1016/j.bbrc.2004.10.012

Notas: ISI