Solution studies to probe active enzymes (cyclophilin-A) and design novel therapeutics (interleukin-8)
EMSL Project ID
46898
Abstract
Overview: A major but essentially unexploited advantage of nuclear magnetic resonance (NMR) spectroscopy is that this technique can be used to monitor active enzymes in solution in order to obtain the atomic resolution details of catalysis. Our applications of NMR methods to enzymes in both the absence of presence of substrates has allowed us to determine how inherent flexibility (dynamics) is critical to enzyme function, and has provided a new paradigm, partially coupled dynamic movements, proposed to be critical to catalytic function of cyclophilin-A. This novel research has far-reaching implications for rational design of biocatalysts, with applications relevant to biomedical and environmental sciences. We seek to obtain preliminary NMR data for two projects in order to submit these as full grant proposals to either the National Science Foundation or the National Institute of Health. Our immediate goals include final data collection (see attached experimental plan) that will allow us to perform the following:
Project #1: Identify the conformational changes that occur DURING CATALYSIS within a cyclophilin-A/substrate complex.
Project #2: Engineer rationally designed interleukin-8 inhibitors through both structure-based and dynamics-based methods.
Project Details
Start Date
2012-01-17
End Date
2013-02-12
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Camilloni C, AB Sahakyan, M Holliday, NG Isern, F Zhang, EZ Eisenmesser, and M Vendruscolo. 2014. "Cyclophilin A catalyzes proline isomerization by an electrostatic handle mechanism." Proceedings of the National Academy of Sciences of the United States of America 111(25):, doi:10.1073/pnas.1404220111
Holliday M, F Zhang, NG Isern, GS Armstrong, and EZ Eisenmesser. 2014. "1H, 13C, and 15N backbone and side chain resonance assignments of thermophilic Geobacillus kaustophilus cyclophilin-A." Biomolecular NMR Assignments 8(1):23-27. doi:10.1007/s12104-012-9445-3