{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,3]],"date-time":"2026-03-03T01:05:32Z","timestamp":1772499932077,"version":"3.50.1"},"reference-count":61,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[2017,8,1]],"date-time":"2017-08-01T00:00:00Z","timestamp":1501545600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"name":"Research council of Norway, FriMedBio","award":["213808"],"award-info":[{"award-number":["213808"]}]},{"name":"Research council of Norway SynkNoyt","award":["218539"],"award-info":[{"award-number":["218539"]}]},{"name":"Research council of Norway, FriMedBio","award":["213808"],"award-info":[{"award-number":["213808"]}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Antimicrob Agents Chemother"],"published-print":{"date-parts":[[2017,8]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Metallo-\u03b2-lactamases (MBLs) threaten the effectiveness of \u03b2-lactam antibiotics, including carbapenems, and are a concern for global public health. \u03b2-Lactam\/\u03b2-lactamase inhibitor combinations active against class A and class D carbapenemases are used, but no clinically useful MBL inhibitor is currently available. Tripoli metallo-\u03b2-lactamase-1 (TMB-1) and TMB-2 are members of MBL subclass B1a, where TMB-2 is an S228P variant of TMB-1. The role of S228P was studied by comparisons of TMB-1 and TMB-2, and E119 was investigated through the construction of site-directed mutants of TMB-1, E119Q, E119S, and E119A (E119Q\/S\/A). All TMB variants were characterized through enzyme kinetic studies. Thermostability and crystallization analyses of TMB-1 were performed. Thiol-based inhibitors were investigated by determining the 50% inhibitory concentrations (IC\n            <jats:sub>50<\/jats:sub>\n            ) and binding using surface plasmon resonance (SPR) for analysis of TMB-1. Thermostability measurements found TMB-1 to be stabilized by high NaCl concentrations. Steady-state enzyme kinetics analyses found substitutions of E119, in particular, substitutions associated with the penicillins, to affect hydrolysis to some extent. TMB-2 with S228P showed slightly reduced catalytic efficiency compared to TMB-1. The IC\n            <jats:sub>50<\/jats:sub>\n            levels of the new thiol-based inhibitors were 0.66 \u03bcM (inhibitor 2a) and 0.62 \u03bcM (inhibitor 2b), and the equilibrium dissociation constant (\n            <jats:italic>\n              K\n              <jats:sub>D<\/jats:sub>\n            <\/jats:italic>\n            ) of inhibitor 2a was 1.6 \u03bcM; thus, both were more potent inhibitors than\n            <jats:sc>l<\/jats:sc>\n            -captopril (IC\n            <jats:sub>50<\/jats:sub>\n            = 47 \u03bcM;\n            <jats:italic>\n              K\n              <jats:sub>D<\/jats:sub>\n            <\/jats:italic>\n            = 25 \u03bcM). The crystal structure of TMB-1 was resolved to 1.75 \u00c5. Modeling of inhibitor 2b in the TMB-1 active site suggested that the presence of the W64 residue results in T-shaped \u03c0-\u03c0 stacking and R224 cation-\u03c0 interactions with the phenyl ring of the inhibitor. In sum, the results suggest that residues 119 and 228 affect the catalytic efficiency of TMB-1 and that inhibitors 2a and 2b are more potent inhibitors for TMB-1 than\n            <jats:sc>l<\/jats:sc>\n            -captopril.\n          <\/jats:p>","DOI":"10.1128\/aac.02602-16","type":"journal-article","created":{"date-parts":[[2017,5,31]],"date-time":"2017-05-31T00:40:34Z","timestamp":1496191234000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":9,"title":["Structural Insights into TMB-1 and the Role of Residues 119 and 228 in Substrate and Inhibitor Binding"],"prefix":"10.1128","volume":"61","author":[{"given":"Susann","family":"Skagseth","sequence":"first","affiliation":[{"name":"The Norwegian Structural Biology Centre (NorStruct), Department of Chemistry, Faculty of Science and Technology, UiT The Arctic University of Norway, Troms\u00f8, Norway"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tony","family":"Christopeit","sequence":"additional","affiliation":[{"name":"The Norwegian Structural Biology Centre (NorStruct), Department of Chemistry, Faculty of Science and Technology, UiT The Arctic University of Norway, Troms\u00f8, Norway"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sundus","family":"Akhter","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Faculty of Science and Technology, UiT The Arctic University of Norway, Troms\u00f8, Norway"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Annette","family":"Bayer","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Faculty of Science and Technology, UiT The Arctic University of Norway, Troms\u00f8, Norway"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"\u00d8rjan","family":"Samuelsen","sequence":"additional","affiliation":[{"name":"Norwegian National 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