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Título:
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High-yield production of a chimeric glycoprotein based on permuted E1 and E2 HCV envelope ectodomains
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Autores:
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Tello, Daniel ;
Rodríguez-Rodríguez, Mar ;
Yélamos, Belén ;
Gómez-Gutiérrez, Julián ;
Peterson, Darrell L. ;
Gavilanes, Francisco
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Tipo de documento:
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texto impreso
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Editorial:
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Elsevier Science BV, 2015-03-01
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Dimensiones:
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application/pdf
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Nota general:
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info:eu-repo/semantics/openAccess
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Idiomas:
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Palabras clave:
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Estado = Publicado
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Materia = Ciencias: Química: Bioquímica
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Tipo = Artículo
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Resumen:
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In this report it is described for the first time the expression and purification of large quantities of a oluble and correctly folded chimeric recombinant protein, E2661E1340, containing the permuted Hepatitis C virus (HCV) glycoprotein ectodomains E1 (amino acids 192-340) and E2 (amino acids 384-661). Using the baculovirus/insect cell expression system, 8mg of secreted protein were purified from 1L of culture media, a yield 4 times higher than the described for its counterpart E1341E2661. This permuted chimeric protein is glycosylated and possesses a high tendency to self-associate. The fluorescence emission spectrum indicates that Trp residues occupy a relatively low hydrophobic environment. The secondary structure was determined by deconvolution of the far-UV circular dichroism spectrum yielding 13% ?-helix structure, 49% extended structure and 38% non-ordered structure. E2661E1340 binds to antibodies present in human sera from HCV-positive patients, a binding that is blocked at different levels by a rabbit anti-E2661 antibody. All these structural and antigenic features of E2661E1340 are very similar to those described for E1340E2661, Thus, this high-yield isolated chimeric protein may be a valuable tool to study the first steps of the HCV infection.
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En línea:
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https://eprints.ucm.es/33660/1/J.%20Virol.%20Meth.%20213%2C%2038-44%20%282015%29.pdf
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