title: Mechanisms leading to an oriented immobilization of recombinant proteins derived from the P24 capsid of HIV-1 onto copolymers.
abstract: To investigate the mechanism leading to an oriented immobilization of recombinant proteins onto synthetic copolymers, five genetically modified HIV-1 p24 capsid proteins (RH24, RH24A4K2, RH24R6, RH24R4K2, and RH24K6) were tested for their efficiency to covalently bind to maleic anhydride-alt-methyl vinyl ether (MAMVE) and N-vinyl pyrrolidone-alt-maleic anhydride (NVPMA) copolymers. These proteins contain, at their C-termini, tags differing in cationic and/or reactive amino acids density. We demonstrated that an increase of the charge and amine density in the tag enhances the coupling yield, the most efficient tag being a six lysine one. The reactivity of the proteins depends directly on the reactivity of the tag, and this led us to conclude that the tag was the site where the covalent grafting with the polymer occurred. Thus, design of such tags provides a new efficient and versatile method allowing oriented immobilization of recombinant proteins onto copolymers.
authors.affiliation: Unité Mixte UMR 2142, CNRS-bioMérieux, ENS-Lyon, 46, allée d'Italie, 69364 Lyon Cédex 07, France.
authors.forename: L
authors.initials: L
authors.lastname: Allard
authors.forename: V
authors.initials: V
authors.lastname: Cheynet
authors.forename: G
authors.initials: G
authors.lastname: Oriol
authors.forename: L
authors.initials: L
authors.lastname: Véron
authors.forename: F
authors.initials: F
authors.lastname: Merlier
authors.forename: G
authors.initials: G
authors.lastname: Scrémin
authors.forename: B
authors.initials: B
authors.lastname: Mandrand
authors.forename: T
authors.initials: T
authors.lastname: Delair
authors.forename: F
authors.initials: F
authors.lastname: Mallet
mesh_terms: D000595:Amino Acid Sequence; D003432:Cross-Linking Reagents / Q000737:chemistry; D016655:HIV Core Protein p24 / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D008299:Maleic Anhydrides / Q000737:chemistry; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry; D011108:Polymers / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D016415:Sequence Alignment
chemical_list: D003432:Cross-Linking Reagents; D016655:HIV Core Protein p24; D008299:Maleic Anhydrides; D009842:Oligopeptides; D011108:Polymers; D011994:Recombinant Proteins