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High-throughput disulfide and FRET scanning to reveal protein conformational ensembles in vitro and in vivo.

Award Number: R00GM141459
ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 09/10/2021
PERIOD OF PERFORMANCE END DATE: 03/31/2027

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High-throughput disulfide and FRET scanning to reveal protein conformational ensembles in vitro and in vivo. - PROJECT SUMMARY Research Strategy Two of the biggest challenges in molecular biomedical research are (1) the lack of technologies for detailed investigation of protein conformations in the cell or the native environment, and (2) a dearth of data on protein conformational dynamics for large proteins and across time scales. This proposal addresses both these grand challenges head-on by developing two closely related technologies: high-throughput disulfide scanning, which will bridge the gap between in vitro and in vivo structural biology by revealing protein structures in both contexts, and single-molecule FRET scanning, which will reveal both average conformation and pairwise distance fluctuations across thousands of residue pairs within a protein in a massively parallel format. I will not only develop these powerful techniques, but also apply them to key biomedical questions: the role of conformational plasticity in the function of chaperones; the role of conformational dynamics in antibiotic susceptibility of bacterial transpeptidases; phenotypic effects of kinetically trapped and non-native protein conformations; and structural basis of genotype-phenotype relationships. Candidate and Environment I am a highly trained experimental protein biochemist and biophysicist with extensive experience investigating the roles of disulfide bonds and redox chemistry in protein structure, stability, misfolding, aggregation, and the pathogenesis of disease. By carrying out the proposed research, I will acquire crucial technical expertise in emerging protein sequencing technologies and computational modeling and data analysis algorithms, as well as a deeper knowledge of statistical physics, microbial physiology, and experimental evolution. Harvard University and the greater Boston-area academic community provides a stellar environment for academic professional development. A group of world-leading experts has been recruited as the Advisory Committee. The primary mentor has a proven track record of training future faculty members at top research-intensive institutions. I will take full advantage of career-development courses, workshops on inclusive teaching and building communities of belonging, seminars on leading a research group, guidance and practice in mentorship, and a formal refresher on responsible conduct of research. I have maintained a high level of productivity throughout my research career (14 publications in 8 years, starting as an undergraduate), and the work arising from this award will be widely disseminated and shared.


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: 2026 ( Subtotal = $249,000 )
2026	2026	THE RESEARCH FOUNDATION FOR THE STATE UNIVERSITY OF NEW YORK	W5510 FRANKS MELVILLE MEMORIAL LIBRARY	STONY BROOK	NY	11794	SUFFOLK	USA	Biomedical Research and Research Training	000	5	4/2/2026	NON-COMPETING CONTINUATION	$249,000
														Subtotal = $249,000

Issue Date FY: 2025 ( Subtotal = $249,000 )
2025	2025	THE RESEARCH FOUNDATION FOR THE STATE UNIVERSITY OF NEW YORK	W5510 FRANKS MELVILLE MEMORIAL LIBRARY	STONY BROOK	NY	11794	SUFFOLK	USA	Biomedical Research and Research Training	000	4	4/11/2025	NON-COMPETING CONTINUATION	$249,000
														Subtotal = $249,000

Issue Date FY: 2024 ( Subtotal = $255,552 )
2024	2024	THE RESEARCH FOUNDATION FOR THE STATE UNIVERSITY OF NEW YORK	W5510 FRANKS MELVILLE MEMORIAL LIBRARY	STONY BROOK	NY	11794	SUFFOLK	USA	Biomedical Research and Research Training	001	3	6/17/2024	EXTENSION WITH OR WITHOUT FUNDS	$6,552
2024	2024	THE RESEARCH FOUNDATION FOR THE STATE UNIVERSITY OF NEW YORK	W5510 FRANKS MELVILLE MEMORIAL LIBRARY	STONY BROOK	NY	11794	SUFFOLK	USA	Biomedical Research and Research Training	000	3	3/21/2024	EXTENSION WITH OR WITHOUT FUNDS	$249,000
														Subtotal = $255,552

Grand Total All Awards = $753,552

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High-throughput disulfide and FRET scanning to reveal protein conformational ensembles in vitro and in vivo.

Award Number: R00GM141459

ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

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

PERIOD OF PERFORMANCE START DATE: 09/10/2021

PERIOD OF PERFORMANCE END DATE: 03/31/2027

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