NMR Studies of the Dynamics of Disordered and Folded States: Isolated and Recombined Fragments from E. coli Thioredoxin.
EMSL Project ID
1976
Abstract
This cooperative project brings together the experience of Dr. Tasayco to design, engineer, generate protein fragments and conduct complementary biophysical experiments to compare the folding of Trx against the analogous folding/binding of its complementary fragments, the expertise of Dr. Gary Daughdrill to carry out NMR relaxation studies of flexible proteins and their corresponding well-structured complexes, and the expertise of Dr. David Lowry to elucidate the mechanism of backbone dynamics. This cooperative proposal has the following aims: 1. To correlate the degree of solvation and flexibility in partially folded Trx fragments. We know from previous protein folding studies that the side chains of hydrophobic residues in the unfolded state are solvent accessible and that this accessibility is significantly reduced in the folded state. However, the way in which the degree of solvation varies with flexibility as folding occurs has not been investigated. We expect a comparison of these two parameters will lead to new insight into the folding process. 2. To correlate the rigidity of the interface between folded, complementary Trx fragments with the same region in native Trx. The mechanism of interface formation in folded proteins remains unclear. This investigation will help us to determine the importance of long range interactions for the folding process. To complete this project we anticipate needing 2 months of 600 MHz instrument time and 2 weeks of 750/800 MHz instrument time.
Project Details
Project type
Capability Research
Start Date
2000-11-15
End Date
2001-03-06
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members