title: Soluble M3 proteins of murine gammaherpesviruses 68 and 72 expressed in Escherichia coli: analysis of chemokine-binding properties.
abstract: M3 protein of murine gammaherpesvirus 68 (MHV-68) was identified as a viral chemokine-binding protein 3 (vCKBP-3) capable to bind a broad spectrum of chemokines and their receptors. During both acute and latent infection MHV-68 M3 protein provides a selective advantage for the virus by inhibiting the antiviral and inflammatory response. A unique mutation Asp307Gly was identified in the M3 protein of murine gammaherpesvirus 72 (MHV-72), localized near chemokine-binding domain. Study on chemokine-binding properties of MHV-72 M3 protein purified from medium of infected cells implied reduced binding to some chemokines when compared to MHV-68 M3 protein. It was suggested that the mutation in the M3 protein might be involved in the attenuation of immune response to infection with MHV-72. Recently, Escherichia coli cells were used to prepare native recombinant M3 proteins of murine gammaherpesviruses 68 and 72 (Pančík et al., 2013). In this study, we assessed the chemokine-binding properties of three M3 proteins prepared in E. coli Rosetta-gami 2 (DE3) cells, the full length M3 protein of both MHV-68 and MHV-72 and MHV-68 M3 protein truncated in the signal sequence (the first 24 aa). They all displayed binding activity to human chemokines CCL5 (RANTES), CXCL8 (IL-8), and CCL3 (MIP-1α). The truncated MHV-68 M3 protein had more than twenty times reduced binding activity to CCL5, but only about five and three times reduced binding to CXCL8 and CCL3 when compared to its full length counterpart. Binding of the full length MHV-72 M3 protein to all chemokines was reduced when compared to MHV-68 M3 protein. Its binding to CCL5 and CCL3 was reduced over ten and seven times. However, its binding to CXCL8 was only slightly reduced (64.8 vs 91.8%). These data implied the significance of the signal sequence and also of a single mutation (at aa 307) for efficient M3 protein binding to some chemokines.
authors.forename: R
authors.initials: R
authors.lastname: Matúšková
authors.forename: P
authors.initials: P
authors.lastname: Pančík
authors.forename: I
authors.initials: I
authors.lastname: Štibrániová
authors.forename: P
authors.initials: P
authors.lastname: Belvončíková
authors.forename: I
authors.initials: I
authors.lastname: Režuchová
authors.forename: M
authors.initials: M
authors.lastname: Kúdelová
mesh_terms: D000818:Animals; D018925:Chemokines / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D018142:Gammaherpesvirinae / Q000145:classification / Q000235:genetics / Q000276:immunology*; D015870:Gene Expression; D006566:Herpesviridae Infections / Q000276:immunology / Q000821:virology*; D006801:Humans; D051379:Mice; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
chemical_list: D018925:Chemokines; C118862:M3 protein, Murine gammaherpesvirus; D011994:Recombinant Proteins; D014764:Viral Proteins
keywords: murine gammaherpesvirus; chemokine-binding protein; M3 protein; signal sequence.