title: In vitro assembly into virus-like particles is an intrinsic quality of Pichia pastoris derived HCV core protein.
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 35nm. 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.
authors.affiliation: Hepatitis C Department, Center for Genetic Engineering and Biotechnology, P.O. Box 6162, C.P. 10600, Cuba. nelson.acosta@cigb.edu.cu
authors.forename: Nelson
authors.initials: N
authors.lastname: Acosta-Rivero
authors.forename: Armando
authors.initials: A
authors.lastname: Rodriguez
authors.forename: Alexis
authors.initials: A
authors.lastname: Musacchio
authors.forename: Viviana
authors.initials: V
authors.lastname: Falcón
authors.forename: Viana M
authors.initials: VM
authors.lastname: Suarez
authors.forename: Gillian
authors.initials: G
authors.lastname: Martinez
authors.forename: Ivis
authors.initials: I
authors.lastname: Guerra
authors.forename: Dalila
authors.initials: D
authors.lastname: Paz-Lago
authors.forename: Yanelys
authors.initials: Y
authors.lastname: Morera
authors.forename: María C
authors.initials: MC
authors.lastname: de la Rosa
authors.forename: Juan
authors.initials: J
authors.lastname: Morales-Grillo
authors.forename: Santiago
authors.initials: S
authors.lastname: Dueñas-Carrera
mesh_terms: D002213:Capsid / Q000737:chemistry / Q000648:ultrastructure; D006801:Humans; D010316:Particle Size; D010843:Pichia / Q000737:chemistry*; D011487:Protein Conformation; D014758:Viral Core Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D014758:Viral Core Proteins; C069130:nucleocapsid protein, Hepatitis C virus