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Understanding the Functional Roles of Newly Identified Serine ‘Orphan’ Proteases and Two Chymotrypsins in the Aedes aegypti Midgut

Award Number: SC3GM116681
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: 02/22/2016
PERIOD OF PERFORMANCE END DATE: 03/31/2024

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Understanding the Functional Roles of Newly Identified Serine ‘Orphan’ Proteases and Two Chymotrypsins in the Aedes aegypti Midgut - Understanding the Functional Roles of Newly Identified Serine `Orphan' Proteases and Two Chymotrypsins in the Aedes aegypti Midgut Project Summary/Abstract The female Aedes aegypti mosquito completely relies on human blood as its main source of nutrients. A single female Ae. aegypti mosquito can lay up to 200 eggs from a single blood meal, and the digestion process is highly dependent on midgut proteolytic enzymes. In vivo studies of four abundant midgut proteases have revealed the importance of only three on maximal fecundity. However, RNAi knockdown studies of all three at once did not have an overall synergistic effect indicating the involvement of other midgut proteases. Indeed, mRNA transcripts of five new serine 'orphan' proteases have been identified, but their physiological roles in this process have not been studied in vivo or in vitro making it unknown how much they contribute to overall fecundity. In addition, the exact role in digestion and overall fecundity of two chymotrypsin-like proteases remains unclear. We hypothesize that the orphan proteolytic and the chymotrypsin-like enzymes play important functional roles in the blood meal digestion process. Because the functions of these proteases are novel in mosquitoes, they will bear broadly on the study of a crucial model organism in vector biology. This proposal will focus on elucidating the physiological roles of these serine proteases using RNAi technology, in vivo and in vitro midgut proteolytic activity assays, fecundity studies (Aim 1), recombinant protein expression and purification for in vitro kinetic and substrate specificity assays, as well as mutational and structural determination using crystallography (Aim 2). Overall, the work will lead to complete understanding of the role and specificity of all known mosquito midgut proteases in the blood meal digestion process and may lead to the development of a potential novel mosquito (vector) control strategy. This is crucial because the mosquito is able to transmit several viral pathogens during blood feeding, and if blood meal digestion is interrupted, the mosquito will not lay eggs minimizing the mosquito population, and in turn minimize pathogen transmission. Furthermore, the information gained from this work may be applied to other vector mosquito species and other blood feeding insect vectors that heavily rely on midgut proteolytic enzymes for digestion.


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 = -$86,309 )
2024	2023	SAN JOSE STATE UNIVERSITY RESEARCH FOUNDATION	210 N 4TH ST FL 4	SAN JOSE	CA	95112	SANTA CLARA	USA	Biomedical Research and Research Training	000	8	8/28/2024	NON-COMPETING CONTINUATION	-$86,309
														Subtotal = -$86,309

Issue Date FY: 2023 ( Subtotal = $109,875 )
2023	2023	SAN JOSE STATE UNIVERSITY RESEARCH FOUNDATION	210 4TH ST 3RD FL	SAN JOSE	CA	95112	SANTA CLARA	USA	Biomedical Research and Research Training	000	8	3/31/2023	NON-COMPETING CONTINUATION	$109,875
														Subtotal = $109,875

Issue Date FY: 2022 ( Subtotal = $109,875 )
2022	2022	SAN JOSE STATE UNIVERSITY RESEARCH FOUNDATION	210 4TH ST 3RD FL	SAN JOSE	CA	95112	SANTA CLARA	USA	Biomedical Research and Research Training	001	7	6/27/2022	NON-COMPETING CONTINUATION	$10,987
2022	2022	SAN JOSE STATE UNIVERSITY RESEARCH FOUNDATION	210 4TH ST 3RD FL	SAN JOSE	CA	95112	SANTA CLARA	USA	Biomedical Research and Research Training	000	7	3/29/2022	NON-COMPETING CONTINUATION	$98,888
														Subtotal = $109,875

Issue Date FY: 2021 ( Subtotal = $109,875 )
2021	2021	SAN JOSE STATE UNIVERSITY RESEARCH FOUNDATION	210 4TH ST 3RD FL	SAN JOSE	CA	95112	SANTA CLARA	USA	Biomedical Research and Research Training	000	6	3/24/2021	NON-COMPETING CONTINUATION	$109,875
														Subtotal = $109,875

Issue Date FY: 2020 ( Subtotal = $100,414 )
2020	2020	SAN JOSE STATE UNIVERSITY RESEARCH FOUNDATION	210 FOURTH ST 4TH FL	SAN JOSE	CA	95112	SANTA CLARA	USA	Biomedical Research and Research Training	001	5	4/3/2020	COMPETING CONTINUATION	$100,414
2020	2020	SAN JOSE STATE UNIVERSITY RESEARCH FOUNDATION	210 FOURTH ST 4TH FL	SAN JOSE	CA	95112	SANTA CLARA	USA	Biomedical Research and Research Training	003	5	5/22/2020	COMPETING CONTINUATION	$83,065
2020	2019	SAN JOSE STATE UNIVERSITY RESEARCH FOUNDATION	210 FOURTH ST 4TH FL	SAN JOSE	CA	95112	SANTA CLARA	USA	Biomedical Research and Research Training	002	4	5/22/2020	NON-COMPETING CONTINUATION	-$83,065
2020	2019	SAN JOSE STATE UNIVERSITY RESEARCH FOUNDATION	210 FOURTH ST 4TH FL	SAN JOSE	CA	95112	SANTA CLARA	USA	Biomedical Research and Research Training	000	4	3/31/2020	NON-COMPETING CONTINUATION	$0
														Subtotal = $100,414

Issue Date FY: 2019 ( Subtotal = $108,375 )
2019	2019	SAN JOSE STATE UNIVERSITY RESEARCH FOUNDATION	210 FOURTH ST 4TH FL	SAN JOSE	CA	95112	SANTA CLARA	USA	Biomedical Research and Research Training	000	4	1/31/2019	NON-COMPETING CONTINUATION	$108,375
														Subtotal = $108,375

Issue Date FY: 2018 ( Subtotal = $108,375 )
2018	2018	SAN JOSE STATE UNIVERSITY RESEARCH FOUNDATION	210 FOURTH ST 4TH FL	SAN JOSE	CA	95112	SANTA CLARA	USA	Biomedical Research and Research Training	001	3	5/11/2018	NON-COMPETING CONTINUATION	$10,838
2018	2018	SAN JOSE STATE UNIVERSITY RESEARCH FOUNDATION	210 FOURTH ST 4TH FL	SAN JOSE	CA	95112	SANTA CLARA	USA	Biomedical Research and Research Training	000	3	12/22/2017	NON-COMPETING CONTINUATION	$97,537
														Subtotal = $108,375

Issue Date FY: 2017 ( Subtotal = $108,375 )
2017	2017	SAN JOSE STATE UNIVERSITY RESEARCH FOUNDATION	210 FOURTH ST 4TH FL	SAN JOSE	CA	95112	SANTA CLARA	USA	Biomedical Research and Research Training	001	2	6/20/2017	NON-COMPETING CONTINUATION	$10,838
2017	2017	SAN JOSE STATE UNIVERSITY RESEARCH FOUNDATION	210 FOURTH ST 4TH FL	SAN JOSE	CA	95112	SANTA CLARA	USA	Biomedical Research and Research Training	000	2	1/13/2017	NON-COMPETING CONTINUATION	$97,537
														Subtotal = $108,375

Issue Date FY: 2016 ( Subtotal = $91,768 )
2016	2016	SAN JOSE STATE UNIVERSITY FOUNDATION	P.O. BOX 720130	SAN JOSE	CA	95172	SANTA CLARA	USA	Biomedical Research and Research Training	000	1	2/22/2016	NEW	$91,768
														Subtotal = $91,768

Grand Total All Awards = $760,623

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Understanding the Functional Roles of Newly Identified Serine ‘Orphan’ Proteases and Two Chymotrypsins in the Aedes aegypti Midgut

Award Number: SC3GM116681

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: 02/22/2016

PERIOD OF PERFORMANCE END DATE: 03/31/2024

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