title: Structure of the c-Src-SH3 domain in complex with a proline-rich motif of NS5A protein from the hepatitis C virus.
abstract: The non-structural hepatitis C virus proteins NS5A and NS5B form a complex through interaction with the SH2 and SH3 domains of the non-receptor Src tyrosine kinase, which seems essential for viral replication. We have crystallized the complex between the SH3 domain of the c-Src tyrosine kinase and the C-terminal proline rich motif of the NS5A protein (A349PPIPPPRRKR359). Crystals obtained at neutral pH belong to the space group I41, with a single molecule of the SH3/NS5A complex at the asymmetric unit. The NS5A peptide is bound in a reverse orientation (class II) and the comparison of this structure with those of the high affinity synthetic peptides APP12 and VSL12 shows some important differences at the salt bridge that drives the peptide orientation. Further conformational changes in residues placed apart from the binding site also seem to play an important role in the binding orientation of this peptide. Our results show the interaction of the SH3 domain of the c-Src tyrosine kinase with a proline rich motif in the NS5A protein and point to their potential interaction in vivo.
authors.affiliation: Department of Chemistry and Physics, University of Almería, Agrifood Campus of International Excellence (ceiA3), Carretera de Sacramento s/n, Almería 04120, Spain.
authors.forename: Julio
authors.initials: J
authors.lastname: Bacarizo
authors.affiliation: Department of Physical Chemistry, University of Granada, Avda. de Fuentenueva s/n, Granada 18071, Spain.
authors.forename: Sergio
authors.initials: S
authors.lastname: Martínez-Rodríguez
authors.affiliation: Department of Chemistry and Physics, University of Almería, Agrifood Campus of International Excellence (ceiA3), Carretera de Sacramento s/n, Almería 04120, Spain. Electronic address: acamara@ual.es.
authors.forename: Ana
authors.initials: A
authors.lastname: Cámara-Artigas
mesh_terms: D000818:Animals; D000081247:CSK Tyrosine-Protein Kinase; D002645:Chickens; D003001:Cloning, Molecular; D003460:Crystallization; D016174:Hepacivirus / Q000737:chemistry*; D008958:Models, Molecular; D046912:Multiprotein Complexes / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D017361:Viral Nonstructural Proteins / Q000737:chemistry*; D019061:src-Family Kinases / Q000737:chemistry*; D012324:RNA-Dependent RNA Polymerase
chemical_list: D000081247:CSK Tyrosine-Protein Kinase; D046912:Multiprotein Complexes; D017361:Viral Nonstructural Proteins; D019061:src-Family Kinases; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase
keywords: Hepatitis C virus; NS5A; Proline rich motifs; SH3 domain; X-ray structure; c-Src tyrosine kinase