title: Processing of the Hepatitis C virus precursor protein expressed in the methylotrophic yeast Pichia pastoris.
abstract: The expression and processing of the Hepatitis C virus core protein (HCcAg) were analyzed in the methylotrophic yeast Pichia pastoris. Two proteins with 21 (p21) and 23 kDa (p23) were detected in immunoblot with a serum from a chronic carrier patient, as the major products for HCcAg. Both proteins, p21 and p23, produced by proteolytic processing in P. partoris, share the same N-terminal region and reacted with a monoclonal antibody towards the first 35 amino acids of HCcAg. The proteolytic processing of the recombinant polypeptide, having the HCcAg and the first 148 aa of E1 protein, was also confirmed by immunoblot analysis using mAbs with HCcAg and E1 specificities, respectively. The 32 kDa glycosilated E1 protein was then immuno-identified, as well as the processed HCcAg. These data demonstrated the usefulness of P. pastoris, as expression system, to study the processing of HCV structural proteins.
authors.affiliation: Division of Vaccines, Center for Genetic Engineering and Biotechnology, Ave. 31 e/158 y 190, P.O. Box 6162, C.P. 10600, La Habana, Cuba.
authors.forename: Nelson
authors.initials: N
authors.lastname: Acosta-Rivero
authors.forename: Alexis
authors.initials: A
authors.lastname: Musacchio
authors.forename: Lazaro
authors.initials: L
authors.lastname: Lorenzo
authors.forename: Catalina
authors.initials: C
authors.lastname: Alvarez
authors.forename: Juan
authors.initials: J
authors.lastname: Morales
mesh_terms: D018936:Hepatitis C Antigens / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D010843:Pichia / Q000235:genetics*; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D014170:Transformation, Genetic; D014758:Viral Core Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*
chemical_list: D018936:Hepatitis C Antigens; D011498:Protein Precursors; D014758:Viral Core Proteins; C069130:nucleocapsid protein, Hepatitis C virus