Our lab works on two main directions: ?rst, the determination of the structural and dynamic basis for the function and assembly of large protein machineries; and second, the determination of the role of internal protein dynamics in regulating protein activity and allosteric interactions. We propose to use NMR spectroscopy, together with other biochemical and biophysical techniques, to determine at the atomic resolution the mechanisms underpinning the function of two important protein families: molecular chaperones and protein kinases. Molecular chaperones are central to maintaining a functional proteome in the cell by rescuing non- native proteins from aggregation and misfolding and assisting with their folding. Our lab reported the ?rst ever high resolution structures of molecular chaperones in complex with unfolded proteins. We will determine the structures of important chaperones such as the Hsp40, Hsp70 and Hsp90 in complex with client proteins. We wish to address how different chaperones engage non-native proteins and how distinct chaperone architectures may alter activity. The Abl kinase holds a prominent place among the over 500 protein kinases encoded by the human genome. Abl mediates its function by participating in a number of biological processes, including actin remodeling, cell adhesion and motility, DNA damage response, and bacterial pathogen response. The Bcr-Abl fusion gene product has constitutive tyrosine kinase activity and causes chronic myeloid leukemia (CML). We will use NMR to provide fascinating, novel information about the regulatory and activation mechanisms of this important kinase. We will study how drug-resistance mutations exert their effect in lowering the drug af?nity for Abl.

Public Health Relevance

Molecular chaperones rescue non-native proteins from aggregation and misfolding and are thus central to maintaining a functional proteome in the cell. Abl fusion proteins (such as Bcr-Abl) are deregulated and cause chronic myeloid leukemia (CML). Both protein families are targets for therapeutic intervention.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Unknown (R35)
Project #
5R35GM122462-03
Application #
9473788
Study Section
Special Emphasis Panel (ZGM1)
Program Officer
Wehrle, Janna P
Project Start
2017-05-01
Project End
2022-04-30
Budget Start
2018-05-01
Budget End
2019-04-30
Support Year
3
Fiscal Year
2018
Total Cost
Indirect Cost
Name
St. Jude Children's Research Hospital
Department
Type
DUNS #
067717892
City
Memphis
State
TN
Country
United States
Zip Code
38105
Saleh, Tamjeed; Rossi, Paolo; Kalodimos, Charalampos G (2017) Atomic view of the energy landscape in the allosteric regulation of Abl kinase. Nat Struct Mol Biol 24:893-901
Monneau, Yoan R; Rossi, Paolo; Bhaumik, Anusarka et al. (2017) Automatic methyl assignment in large proteins by the MAGIC algorithm. J Biomol NMR 69:215-227