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. Protein kinases are a diverse set of regulatory enzymes responsible for signaling in a majority of metabolic pathways and cell behaviors. Kinases control signaling events within the cell through phosphorylation of substrate proteins. Identification of a given kinases'substrate proteins and their specific phosphorylation sites is essential to understanding kinases mediated signaling events. However, there is no general or trivial method for such identifications. We are investigating a method of elucidating kinase phosphorylation sites using small molecule inhibitors of a specific kinase, Nek2, as well as an inhibitor resistant mutant of Nek2 kinase. The method involves over-expressing the kinase of interest (Nek2) in human cells with an epitope tag that can be used to purify Nek2 and associated proteins from these cells. Mass Spec analysis can then be used to identify phosphorylation sites on these purified proteins. By making a quantitative comparison between samples treated with out Nek2 inhibitor and samples expressing the inhibitor resistant Nek2 mutant we can determine phosphorylation sites specifically linked to Nek2 kinase activity.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR001614-29
Application #
8363809
Study Section
Special Emphasis Panel (ZRG1-BCMB-M (40))
Project Start
2011-06-01
Project End
2012-05-31
Budget Start
2011-06-01
Budget End
2012-05-31
Support Year
29
Fiscal Year
2011
Total Cost
$17
Indirect Cost
Name
University of California San Francisco
Department
Pharmacology
Type
Schools of Pharmacy
DUNS #
094878337
City
San Francisco
State
CA
Country
United States
Zip Code
94143
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