Design of N-substituted peptomer ligands for EVH1 domains.
Ena/VASP proteins are implicated in cytoskeletal reorganization during actin-dependent motility processes. Recruitment to subcellular sites of actin polymerization is mediated by the highly conserved N-terminal EVH1 domain, which interacts with target proteins containing proline-rich motifs. The VASP EVH1 domain specifically binds peptides with the consensus motif FPPPP present in all its binding partners, including the Listerial ActA protein. Previous studies have shown that the Phe and first and final Pro residues are highly conserved and cannot be substituted with any other natural amino acid without significant loss of binding affinity. We have incorporated peptoid building blocks (sarcosine derived, non-natural amino acids) into the peptide SFEFPPPPTEDEL from the Listerial ActA protein and were able to substitute the most highly conserved residues of this motif while maintaining binding to the VASP EVH1 domain with affinities in the range of 45-180 microm. We then used NMR chemical shift perturbations to locate specific domain residues involved in particular interactions. These studies may open up the way for designing selective modulators of VASP function for biological studies and for the development of novel therapeutics for diseases involving pathologically altered cell adhesion or cell motility.
Zimmermann
Jürgen
J
Forschungsinstitut für Molekulare Pharmakologie, Robert-Rössle-Strasse 10, 13125 Berlin, Germany.
Kühne
Ronald
R
Volkmer-Engert
Rudolf
R
Jarchau
Thomas
T
Walter
Ulrich
U
Oschkinat
Hartmut
H
Ball
Linda J
LJ
D000199:Actins / Q000378:metabolism; D020816:Amino Acid Motifs; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000737:chemistry; D002465:Cell Movement; D002482:Cellulose / Q000737:chemistry; D003599:Cytoskeleton / Q000378:metabolism; D015195:Drug Design; D007700:Kinetics; D008024:Ligands; D008089:Listeria monocytogenes / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism; D008840:Microfilament Proteins; D008956:Models, Chemical; D008958:Models, Molecular; D010455:Peptides / Q000737:chemistry; D010750:Phosphoproteins / Q000737:chemistry; D011392:Proline / Q000737:chemistry; D017434:Protein Structure, Tertiary; D000098662:Vasodilator-Stimulated Phosphoprotein
D000199:Actins; D001426:Bacterial Proteins; D015815:Cell Adhesion Molecules; D008024:Ligands; D008565:Membrane Proteins; D008840:Microfilament Proteins; D010455:Peptides; D010750:Phosphoproteins; D000098662:Vasodilator-Stimulated Phosphoprotein; C072980:actA protein, Listeria monocytogenes; D002482:Cellulose; D011392:Proline