The solution structure of pGolemi, a high affinity Mena EVH1 binding miniature protein, suggests explanations for paralog-specific binding to Ena/VASP homology (EVH) 1 domains.
Ena/VASP homology 1 (EVH1) domains are polyproline binding domains that are present in a wide range of adaptor proteins, among them Ena/VASP proteins involved in actin remodeling and axonal guidance. The interaction of ActA, a transmembrane protein from the food-borne pathogen Listeria monocytogenes, with EVH1 domains has been shown to be crucial for recruitment of the host's actin skeleton and, as a consequence, for the infectivity of this bacterium. We present the structure of a synthetic high-affinity Mena EVH1 ligand, pGolemi, capable of paralog-specific binding, solved by NMR spectroscopy. This peptide shares the common pancreatic peptide fold with its scaffold, avian pancreatic peptide, but shows pivotal differences in the amino-terminus. The interplay of spatial fixation and flexibility appears to be the reason for its high affinity towards Mena EVH1. Combined with earlier investigations, our structural data shed light on the specificity determinants of pGolemi and the importance of additional binding epitopes around the residues Thr74 and Phe32 on EVH1 domains regulating paralog specificity. Our results are expected to facilitate the design of other high-affinity, paralog-specific EVH1 domain ligands, and serve as a fundament for the investigation of the molecular mode of action of EVH1 domains.
Link
Nina M
NM
Medizinische und Strukturelle Biochemie, Zentrum für Medizinische Biotechnologie, Universität Duisburg-Essen, Universitätsstr. 2-5, D-45117 Essen, Germany. nina.link@uni-due.de
Hunke
Cornelia
C
Mueller
Jonathan W
JW
Eichler
Jutta
J
Bayer
Peter
P
D000595:Amino Acid Sequence; D000818:Animals; D030161:Avian Proteins / Q000737:chemistry / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D008024:Ligands; D008089:Listeria monocytogenes / Q000737:chemistry* / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D055232:Proline-Rich Protein Domains; D011485:Protein Binding; D011487:Protein Conformation; D016415:Sequence Alignment; D040681:Structural Homology, Protein
D030161:Avian Proteins; D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; C092744:ENA-VASP proteins; D008024:Ligands; D008565:Membrane Proteins; D010455:Peptides; C072980:actA protein, Listeria monocytogenes