Sunday, October 27, 2024 10/27/2024

Exploring the Dynamics of Prolyl-tRNA Synthetases: Towards Developing a Screening Method for Species-Specific Inhibitors

Award Number: R15GM117510
ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

Group Awards By:

View Award Abstract

Broadly speaking, our research group studies the interplay between enzyme dynamics and catalysis. This supplemental proposal is developed to acquire Isothermal Titration Calorimeter, which will aid in investigating the effects of molecular crowding on enzyme structure-dynamics- functions. The interior of a cell is extremely crowded, which can potentially impact protein properties such as structure, folding, stability, ligand binding, and enzyme catalysis. The crowding effects could mediate through hard/steric/entropic and/or soft/chemical/enthalpic interactions. Our objective is to investigate the exact mechanism of crowding/confinement and their impacts on substrate binding and catalysis. We are studying the multidomain prolyl-tRNA synthetases (ProRSs), which play a vital role in protein synthesis in all living organisms. Earlier, we demonstrated that domain dynamics is central to ProRSs functions, which are impacted by the crowder-induced shift in the conformational ensemble. Currently, the impact of the “uniform crowding environment” (nonprotein-based crowders) and “structured crowding environment” (protein-based crowders) on conformational dynamics and function of Escherichia coli (Ec) ProRS is being probed. The mechanistic implications of crowding and confinement effects are significant as these enzymes are promising anti-microbial drug targets. The conformational change and crowder-enzyme interactions are being probed by fluorescence spectroscopy, molecular dynamics simulations, and small molecule docking studies, while varying the size, shape, chemical nature, and concentration of the crowders. We are examining the role of molecular crowders on the substrate binding using Saturation Transfer Difference - Nuclear Magnetic Resonance, which requires a large quantity of proteins and kinetics parameters are obtained from enzyme kinetics assays using radiolabeled substrates. The isothermal titration calorimetry (ITC) method is a label-free direct measurement of the heat evolved or absorbed during a binding event between interacting molecules and enzyme-catalyzed reactions and requires less sample. ITC experiments provide the thermodynamic profiles, which would enable us to establish the molecular mechanism of crowding and confinement resulting in enzyme's altered function. A detailed understanding of crowding effects on the properties of multidomain proteins like Ec ProRS could open up new possibilities for protein designing and drug discovery. Moreover, the ITC equipment would enable us to provide hands-on experiences to students in biochemistry and biophysical chemistry courses (~25 students/semester) by designing course-embedded research projects on calorimetry and teaching them the ligand binding thermodynamics in drug discovery.


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2024 ( Subtotal = $0 )
2024	2023	UNIVERSITY OF WISCONSIN SYSTEM	105 GARFIELD AVE	EAU CLAIRE	WI	54701	EAU CLAIRE	USA	Biomedical Research and Research Training	001	2	2/14/2024	SUPPLEMENT FOR EXPANSION	$0
2024	2021	UNIVERSITY OF WISCONSIN SYSTEM	105 GARFIELD AVE	EAU CLAIRE	WI	54701	EAU CLAIRE	USA	Biomedical Research and Research Training	000	2	2/14/2024	COMPETING CONTINUATION	$0
														Subtotal = $0

Issue Date FY: 2023 ( Subtotal = $99,900 )
2023	2023	UNIVERSITY OF WISCONSIN SYSTEM	105 GARFIELD AVE	EAU CLAIRE	WI	54701	EAU CLAIRE	USA	Biomedical Research and Research Training	000	2	5/23/2023	SUPPLEMENT FOR EXPANSION	$99,900
														Subtotal = $99,900

Issue Date FY: 2021 ( Subtotal = $397,331 )
2021	2021	UNIVERSITY OF WISCONSIN SYSTEM	105 GARFIELD AVE	EAU CLAIRE	WI	54701	EAU CLAIRE	USA	Biomedical Research and Research Training	002	2	9/29/2021	COMPETING CONTINUATION	$952
2021	2021	UNIVERSITY OF WISCONSIN SYSTEM	105 GARFIELD AVE	EAU CLAIRE	WI	54701	EAU CLAIRE	USA	Biomedical Research and Research Training	000	2	3/17/2021	COMPETING CONTINUATION	$397,331
2021	2016	UNIVERSITY OF WISCONSIN SYSTEM	105 GARFIELD AVE	EAU CLAIRE	WI	54701	EAU CLAIRE	USA	Biomedical Research and Research Training	001	1	9/29/2021	NEW	-$952
														Subtotal = $397,331

Issue Date FY: 2020 ( Subtotal = $0 )
2020	2016	UNIVERSITY OF WISCONSIN SYSTEM	105 GARFIELD AVE	EAU CLAIRE	WI	54701	EAU CLAIRE	USA	Biomedical Research and Research Training	000	1	1/16/2020	NEW	$0
														Subtotal = $0

Issue Date FY: 2016 ( Subtotal = $380,899 )
2016	2016	UNIVERSITY OF WISCONSIN, EAU CLAIRE	105 GARFIELD AVENUE	EAU CLAIRE	WI	54701	EAU CLAIRE	USA	Biomedical Research and Research Training	000	1	1/11/2016	NEW	$336,991
2016	2016	UNIVERSITY OF WISCONSIN, EAU CLAIRE	105 GARFIELD AVENUE	EAU CLAIRE	WI	54701	EAU CLAIRE	USA	Biomedical Research and Research Training	001	1	4/14/2016	NEW	$43,908
														Subtotal = $380,899

Grand Total All Awards = $878,130

Top

All Categories

About

Search

Reports

Data Submission

Award Information

Exploring the Dynamics of Prolyl-tRNA Synthetases: Towards Developing a Screening Method for Species-Specific Inhibitors

Award Number: R15GM117510

ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

Federal Websites

Department of Health & Human Services

HHS Operating Divisions

HHS Staff Divisions

Download A Document Viewer