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...

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Published in:Biochemistry (Easton) 2002-09, Vol.41 (37), p.11143-11151
Main Authors: Zimmermann, Jürgen, Labudde, Dirk, Jarchau, Thomas, Walter, Ulrich, Oschkinat, Hartmut, Ball, Linda J
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Biopolymers - chemistry
Cell Adhesion Molecules - chemistry
Centrifugation, Density Gradient
Circular Dichroism
DNA-Binding Proteins - chemistry
Dogs
Humans
Hydrogen-Ion Concentration
Mice
Microfilament Proteins
Molecular Sequence Data
Nitrogen Isotopes
Nuclear Magnetic Resonance, Biomolecular - methods
Peptide Fragments - chemistry
Phosphoproteins - chemistry
Protein Structure, Secondary
Protein Structure, Tertiary
Rats
Sequence Homology, Amino Acid
Temperature
Thermodynamics
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Summary: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 15N-HSQC-experiment exhibits only one set of peaks, suggesting a high degree of symmetry for a putative oligomer. Measurements of the rotational correlation time τC of the molecule and analytical ultracentrifugation data show VASP (336−380) to be entirely tetrameric in solution. The secondary structure was confirmed from a 15N-NOESY-HSQC experiment and is completely α-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.
ISSN:0006-2960
1520-4995
DOI:10.1021/bi020379x