Relaxation, equilibrium oligomerization, and molecular symmetry of the VASP (336-380) EVH2 tetramer.
An investigation of the structural and dynamic properties of the C-terminal fragment of the human protein VASP (VASP 336-380) has been performed. Full length VASP has been shown to be tetrameric in solution, and the C-terminal 45 residues of the protein have been suggested to be responsible for the oligomerization. We have expressed and purified a C-terminal fragment of the human VASP protein from residue 336-380. It was found to form a stable domain in its own right. The fragment was shown by CD spectroscopy to form a helical structure, stable under a wide range of temperature and pH conditions. A (15)N-HSQC-experiment exhibits only one set of peaks, suggesting a high degree of symmetry for a putative oligomer. Measurements of the rotational correlation time tau(C) of the molecule and analytical ultracentrifugation data show VASP (336-380) to be entirely tetrameric in solution. The secondary structure was confirmed from a (15)N-NOESY-HSQC experiment and is completely alpha-helical. We conclude that VASP (336-380) forms a tetramer in solution via a coiled coil arrangement and is solely responsible for tetramerization of full-length VASP.
Zimmermann
Jürgen
J
Forschungsinstitut für Molekulare Pharmakologie, Robert-Rössle-Strasse 10, 13125 Berlin, Germany. zimmermann@fmp-berlin.de
Labudde
Dirk
D
Jarchau
Thomas
T
Walter
Ulrich
U
Oschkinat
Hartmut
H
Ball
Linda J
LJ
D000595:Amino Acid Sequence; D000818:Animals; D001704:Biopolymers / Q000737:chemistry; D015815:Cell Adhesion Molecules / Q000737:chemistry*; D002499:Centrifugation, Density Gradient; D002942:Circular Dichroism; D004268:DNA-Binding Proteins / Q000737:chemistry*; D004285:Dogs; D006801:Humans; D006863:Hydrogen-Ion Concentration; D051379:Mice; D008840:Microfilament Proteins; D008969:Molecular Sequence Data; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D010446:Peptide Fragments / Q000737:chemistry*; D010750:Phosphoproteins / Q000737:chemistry*; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D051381:Rats; D017386:Sequence Homology, Amino Acid; D013696:Temperature; D013816:Thermodynamics; D000098662:Vasodilator-Stimulated Phosphoprotein
D001704:Biopolymers; D015815:Cell Adhesion Molecules; D004268:DNA-Binding Proteins; C092744:ENA-VASP proteins; D008840:Microfilament Proteins; D009587:Nitrogen Isotopes; D010446:Peptide Fragments; D010750:Phosphoproteins; D000098662:Vasodilator-Stimulated Phosphoprotein