NMR Assignment & Structure of the C-Terminal Domains of Human Villin
EMSL Project ID
17990
Abstract
Proteins are often composed of modular domains and NMR is uniquely suited to exploring the solution structure and dynamics of interactions between moderately sized domains. We are interested in determining the structure and, ultimately, the dynamics of the two, C-terminal domains of human villin. Villin is a multifunctional, modular, F-actin bundling and calcium-sensitive severing protein found in the absorptive epithelium that lines the gut and the proximal tube in the kidney. Villin is essential for the entry of the microbe Shigella flexneri into humans that causes bacillary dysentery that results in over one million deaths per year world wide, mostly in children. We have focused on the C-terminal domains of villin as a model system because the structure of each domain (or their homologs) have been determined, but nothing is known about them as a linked, modular system. The requested NMR experiments will assist in the assignment and structure determination of the sixth gelsolin-like domain of villin (~100 residues), a ~ 30 residue sequence that links the two domains together, and the 76-residue, C-terminal headpiece domain. This 208 amino acid construct is expressed at high levels in 13C/15N media and spectra on our 500 MHz NMR instrument indicate that it is well folded. We are requesting time at higher field to aid our efforts to assign the resonances and determine the structure of this construct. The project is funded by the NIH. We hope to work closely and collaboratively with the staff at EMSL. Our samples are already made and stable so that we are flexible with respect of scheduling. The proposed research should shed much needed light on the interactions and dynamics of domain-domain interactions modular proteins.
Project Details
Project type
Capability Research
Start Date
2006-06-08
End Date
2007-06-12
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Smirnov S, NG Isern, ZG Jiang, DW Hoyt, and CJ Mcknight. 2007. "The Isolated Sixth Gelsolin Repeat and Headpiece Domain of Villin Bundle F-Actin in the Presence of Calcium and Are Linked by a 40-Residue Unstructured Sequence ." Biochemistry 46(25):7488-7496.