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Real-time estimation of antibiotic concentration in a bacterial cell

Award Number: R21AI125873
ORGANIZATION: NATIONAL INSTITUTE OF ALLERGY & INFECTIOUS DISEASES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 06/15/2016
PERIOD OF PERFORMANCE END DATE: 11/30/2018

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Real-time estimation of antibiotic concentration in a bacterial cell - Project Summary A real-time estimation of antibiotic in a bacterium is very crucial for understanding how that molecule is processed inside the cell. This will also help understand the mechanism of resistance. We propose to design a genetically encoded biosensor that can bind to the antibiotic of interest and produce a convenient signal. Using periplasmic binding protein (PBP) family, that is known to bind to diverse set of small molecules as a scaffold, we propose to engineer split PBP, consisting of globular domains, that can sandwich a small molecule of interest, an antibiotic in the present case. Earlier work with PBP used the whole protein with limited success; a single scaffold only gave restricted opportunity to carry mutagenesis and design. We also propose that two domains when undergo ligand mediated binding, will show predictable and large change in interdomain distance and orientation. The spatial change can be exploited for a quantitative FRET increase, which can be correlated to the concentration of the antibiotic. The proposal brings together all aspects of computational protein design, that include, loop modeling, stability and solubility enhancement, protein interface design and finally ligand docking. Our protein engineering approach carefully meshes computational modeling with flow cytomtery assisted high throughput screening to create functional proteins with applications in therapy and diagnostics.


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: 2019 ( Subtotal = -$5,857 )
2019	2017	LOS ALAMOS NATIONAL SECURITY, LLC	335 CENTRAL PARK SQ	LOS ALAMOS	NM	87544	LOS ALAMOS	USA	Allergy and Infectious Diseases Research	000	2	4/25/2019	NON-COMPETING CONTINUATION	-$5,857
														Subtotal = -$5,857

Issue Date FY: 2017 ( Subtotal = $233,160 )
2017	2017	LOS ALAMOS NATIONAL SECURITY, LLC	BIKINI ATOLL RD SM 30	LOS ALAMOS	NM	87545	LOS ALAMOS	USA	Allergy and Infectious Diseases Research	000	2	5/17/2017	NON-COMPETING CONTINUATION	$233,160
														Subtotal = $233,160

Issue Date FY: 2016 ( Subtotal = $291,946 )
2016	2016	LOS ALAMOS NATIONAL SECURITY LLC	4200 WEST JEMEZ ROAD	LOS ALAMOS	NM	87544	LOS ALAMOS	USA	Allergy and Infectious Diseases Research	001	1	6/22/2016	NEW	$0
2016	2016	LOS ALAMOS NATIONAL SECURITY LLC	4200 WEST JEMEZ ROAD	LOS ALAMOS	NM	87544	LOS ALAMOS	USA	Allergy and Infectious Diseases Research	000	1	6/14/2016	NEW	$291,946
														Subtotal = $291,946

Grand Total All Awards = $519,249

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Real-time estimation of antibiotic concentration in a bacterial cell

Award Number: R21AI125873

ORGANIZATION: NATIONAL INSTITUTE OF ALLERGY & INFECTIOUS DISEASES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

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

PERIOD OF PERFORMANCE START DATE: 06/15/2016

PERIOD OF PERFORMANCE END DATE: 11/30/2018

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