Characterizing and controlling the inherent dynamics of cyclophilin-A |
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Authors: | Jennifer Schlegel Geoffrey S Armstrong Jasmina S Redzic Fengli Zhang Elan Zohar Eisenmesser |
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Institution: | 1.Department of Biochemistry and Molecular Genetics, School of Medicine, University of Colorado Denver, Aurora, Colorado 80045;2.Department of Chemistry and Biochemistry, University of Colorado at Boulder, Boulder, Colorado 80309;3.National High Magnetic Field Laboratory, Tallahassee, Florida 32310 |
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Abstract: | With the recent advances in NMR relaxation techniques, protein motions on functionally important timescales can be studied at atomic resolution. Here, we have used NMR-based relaxation experiments at several temperatures and both 600 and 900 MHz to characterize the inherent dynamics of the enzyme cyclophilin-A (CypA). We have discovered multiple chemical exchange processes within the enzyme that form a “dynamic continuum” that spans 20–30 Å comprising active site residues and residues proximal to the active site. By combining mutagenesis with these NMR relaxation techniques, a simple method of counting the dynamically sampled conformations has been developed. Surprisingly, a combination of point mutations has allowed for the specific regulation of many of the exchange processes that occur within CypA, suggesting that the dynamics of an enzyme may be engineered. |
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Keywords: | chemical exchange cyclophilin-A dynamics peptidyl-prolyl isomerase relaxation nuclear magnetic resonances enzymes NMR spectroscopy |
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