{"id":1047,"date":"2018-06-26T17:20:00","date_gmt":"2018-06-26T21:20:00","guid":{"rendered":"https:\/\/research.cbc.osu.edu\/gopalan.5\/?page_id=1047"},"modified":"2019-11-05T15:17:46","modified_gmt":"2019-11-05T20:17:46","slug":"salmonella_metabolism","status":"publish","type":"page","link":"https:\/\/research.cbc.osu.edu\/gopalan.5\/salmonella_metabolism\/","title":{"rendered":"Metabolic Pathways in Salmonella enterica"},"content":{"rendered":"

Metabolic Pathways in Salmonella enterica<\/em><\/strong><\/h2>\n
\n

We are investigating the individual roles of an asparaginase, a kinase, a deglycase, and a putative transcriptional regulator in the utilization of fructose-asparagine (F-Asn), an Amadori product, by Salmonella<\/em>. In particular, we are exploring whether this distinctive metabolic capability may be exploited to design narrow-spectrum anti-Salmonella<\/em> therapeutics.<\/p><\/blockquote>\n

\n

I. NIH R01AI116119 (December 2014 – November 2019)<\/strong><\/p>\n

Title<\/em>
\nSalmonella<\/em>, colonization resistance, and fructose-asparagine<\/p>\n

Narrative<\/em>
\nThe metabolism of fructose-asparagine is required for Salmonella<\/em> to thrive in the intestine, making the acquisition system a novel drug target. We will learn which intestinal organisms compete with Salmonella<\/em> for fructose-asparagine and why they fail, which will facilitate the development of novel probiotics that can prevent or treat Salmonella<\/em> infections. We will also determine the enzymatic mechanisms used by Salmonella<\/em> to grow on fructose-asparagine, which will facilitate high-throughput screens for inhibitors of these enzymes.<\/p>\n

Senior personnel<\/em>
\n\u2022   Brian M.M. Ahmer (OSU, lead PI)
\n\u2022   Edward J. Behrman, Venkat Gopalan, Kelly C. Wrighton, Vicki H. Wysocki (OSU, PIs)<\/p>\n

Lab members involved in this project<\/em>
\n\u2022   Sravya Kovvali (Graduate Student, OSU Microbiology Graduate Program)
\n\u2022   Blake E. Szkoda (Graduate Student, Ohio State Biochemistry Program)<\/p>\n

Methods<\/em>
\n\u2022   In our laboratory:<\/strong> protein overexpression and purification; enzyme kinetic studies; DNA\u2014protein footprinting; fluorescence polarization (FP) binding assays
\n\u2022   In collaboration with other PIs:<\/strong> in vivo<\/em> studies (Ahmer); synthesis of Amadori products (Behrman); protein crystallography (Charles Bell, OSU); protein modeling (Steffen Lindert, OSU); mass spectrometry (Wysocki)<\/p>\n

Recent Publications<\/em>
\n\u2022   Sengupta A, Wu J, Seffernick JT, Sabag-Daigle A, Thomsen N, Chen T-H, Di Capua A, Bell CE, Ahmer BMM, Lindert S, Wysocki VH, and Gopalan V*. (2019) Integrated use of biochemical, native mass spectrometry, computational and genome-editing methods to elucidate the mechanism of a Salmonella<\/em> deglycase. J. Mol. Biol.<\/em>, in press<\/em>.
\n\u2022   Wu J\u00b0, Sabag-Daigle A\u00b0, Borton MA^, Kop LFM^, Szkoda BE^, Deatherage Kaiser BL, Lindemann SR, Renslow RS, Wei S, Nicora CD, Weitz KK, Kim Y-M, Adkins JN, Metz TO, Boyaka P, Gopalan V, Wrighton KC, Wysocki VH*, and Ahmer BMM*. (2018) Salmonella<\/em>-mediated inflammation eliminates competitors for fructose-asparagine in the gut. Infect. Immun.<\/em>, 86<\/strong>: e00945-17. \u00b0joint first authors; ^joint second authors<\/em><\/strong>
\n\u2022   Wu J, Sabag-Daigle A, Metz TO, Deatherage Kaiser BL, Gopalan V, Behrman EJ, Wysocki VH*, and Ahmer BMM*. (2018) Measurement of fructose\u2013asparagine concentrations in human and animal foods. J. Agric. Food Chem.<\/em>, 66<\/strong>: 212-217.
\n\u2022   Sabag-Daigle A, Wu J, Borton MA, Sengupta A, Gopalan V, Wrighton KC, Wysocki VH, and Ahmer BMM*. (2018) Identification of bacterial species that can utilize fructose-asparagine. Appl. Environ. Microbiol.<\/em>, 84<\/strong>: e01957-17.
\n\u2022   Sabag-Daigle A, Sengupta A, Blunk HM, Biswas PK, Cron MC, Bogard AJ, Behrman EJ, Gopalan V, and Ahmer BMM*. (2017) Salmonella<\/em> FraE, an asparaginase homolog, contributes to fructose-asparagine but not asparagine utilization. J. Bacteriol.<\/em>, 199<\/strong>: e00330-17.<\/a>
\n\u2022   Biswas PK, Behrman EJ*, and Gopalan V*. (2017) Characterization of a Salmonella<\/em> sugar kinase essential for utilization of fructose-asparagine. Biochem. Cell Biol.<\/em>, 95<\/strong>: 304-309.
\n\u2022   Sabag-Daigle A, Blunk HM, Sengupta A, Wu J, Bogard AJ, Ali MM, Stahl C, Wysocki VH, Gopalan V, Behrman EJ, and Ahmer BMM*. (2016) A metabolic intermediate of the fructose-asparagine utilization pathway inhibits growth of a Salmonella fraB<\/em> mutant. Sci. Rep.<\/em>, 6<\/strong>: 28117.<\/a> [SI<\/a>]<\/p>\n

\n

II. NIH R01AI140541 (August 2018 – July 2022)<\/strong><\/p>\n

Title<\/em>
\nSalmonella<\/em>-specific therapeutics<\/p>\n

Narrative<\/em>
\nNon-typhoidal salmonellosis is a serious threat in the United States and globally. Because Salmonella<\/em> cells lacking the FraB deglycase of the fructose-asparagine (F-Asn) utilization pathway accumulate a toxic intermediate during F-Asn metabolism, FraB is a valid and exploitable drug target. Our objectives in this proposal are to identify and characterize potent inhibitors of FraB, and provide a foundation for our long-term goal of developing a novel therapeutic for non-typhoidal salmonellosis.<\/p>\n

Senior personnel<\/em>
\n\u2022   Brian M.M. Ahmer (OSU, lead PI)
\n\u2022   Venkat Gopalan (OSU, PI)
\n\u2022   Edward J. Behrman, Charles E. Bell, Steffen Lindert (OSU, co-Is)<\/p>\n

Lab members involved in this project<\/em>
\n\u2022   Sravya Kovvali (Graduate Student, OSU Microbiology Graduate Program)
\n\u2022   Jamison Law (Graduate Student, Ohio State Biochemistry Program)<\/p>\n

Methods<\/em>
\n\u2022   In our laboratory:<\/strong> high-throughput biochemical screening, validation, and characterization of small molecule inhibitors
\n\u2022   In collaboration with other PIs:<\/strong> in vivo<\/em> screening (Ahmer); synthesis of fructose-asparagine derivatives and lead optimization (Behrman); protein crystallography (Bell); in silico<\/em> structure-based virtual screening (Lindert); mass spectrometry (Vicki Wysocki, OSU)<\/p>\n","protected":false},"excerpt":{"rendered":"

Metabolic Pathways in Salmonella enterica We are investigating the individual roles of an asparaginase, a kinase, a deglycase, and a putative transcriptional regulator in the utilization of fructose-asparagine (F-Asn), an Amadori product, by Salmonella. In particular, we are exploring whether this distinctive metabolic capability may be exploited to design narrow-spectrum anti-Salmonella therapeutics. I. NIH R01AI116119 […]<\/p>\n","protected":false},"author":9,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_exactmetrics_skip_tracking":false,"_exactmetrics_sitenote_active":false,"_exactmetrics_sitenote_note":"","_exactmetrics_sitenote_category":0,"footnotes":""},"class_list":["post-1047","page","type-page","status-publish","hentry","post-preview"],"jetpack_shortlink":"https:\/\/wp.me\/P8Lmk8-gT","jetpack_sharing_enabled":true,"_links":{"self":[{"href":"https:\/\/research.cbc.osu.edu\/gopalan.5\/wp-json\/wp\/v2\/pages\/1047","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/research.cbc.osu.edu\/gopalan.5\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/research.cbc.osu.edu\/gopalan.5\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/research.cbc.osu.edu\/gopalan.5\/wp-json\/wp\/v2\/users\/9"}],"replies":[{"embeddable":true,"href":"https:\/\/research.cbc.osu.edu\/gopalan.5\/wp-json\/wp\/v2\/comments?post=1047"}],"version-history":[{"count":27,"href":"https:\/\/research.cbc.osu.edu\/gopalan.5\/wp-json\/wp\/v2\/pages\/1047\/revisions"}],"predecessor-version":[{"id":1705,"href":"https:\/\/research.cbc.osu.edu\/gopalan.5\/wp-json\/wp\/v2\/pages\/1047\/revisions\/1705"}],"wp:attachment":[{"href":"https:\/\/research.cbc.osu.edu\/gopalan.5\/wp-json\/wp\/v2\/media?parent=1047"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}