This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Allosteric communication between remote sites is a crucial element in enzymatic function. Using TROSY-based NMR spectroscopy, we will map long-range allosteric changes along the backbone of the catalytic subunit of cAMP-dependent protein kinase A (PKA) upon binding a nucleotide analogue (AMP-PNP) and the peptide LRRASLG (Kemptide) as substrates. Calculations for the binding constants of the two ligands, as well as for on and off rates will be obtained. Since the enzyme retains activity under our NMR conditions, the structural changes monitored in this study will reflect the interconversion of conformations during turnover. These studies represent a paradigm for how multidomain kinase turnover involves complex dynamics transmitted within its structure. In addition, this is the first report of monitored dynamic changes within an active kinase at atomic resolution.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR002301-26
Application #
8361154
Study Section
Special Emphasis Panel (ZRG1-BCMB-H (40))
Project Start
2011-03-01
Project End
2012-02-29
Budget Start
2011-03-01
Budget End
2012-02-29
Support Year
26
Fiscal Year
2011
Total Cost
$8,702
Indirect Cost
Name
University of Wisconsin Madison
Department
Biochemistry
Type
Schools of Earth Sciences/Natur
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
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