{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T05:24:27Z","timestamp":1791523467399,"version":"4.3.4"},"reference-count":51,"publisher":"Wiley","issue":"8","license":[{"start":{"date-parts":[[2009,1,1]],"date-time":"2009-01-01T00:00:00Z","timestamp":1230768000000},"content-version":"vor","delay-in-days":1614,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2004,8]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Metallo\u2010\u03b2\u2010lactamase L1, secreted by pathogenic <jats:italic>Stenotrophomonas maltophilia<\/jats:italic>, is a dinuclear Zn(II)\u2010containing enzyme that hydrolyzes almost all known penicillins, cephalosporins, and carbapenems. The presence of Zn(II) ions in both metal binding sites is essential for full enzymatic activity; however, the mechanism of physiological metal incorporation is unknown. To probe metal incorporation, L1 was over\u2010expressed in minimal media with (mmL1+Zn) and without (mmL1\u2212Zn) Zn(II) added to the media, and the resulting proteins were purified and characterized. The mmL1+Zn sample was bound by a Q\u2010Sepharose column, exhibited steady\u2010state kinetic properties, bound Zn(II), existed as a tetramer, and yielded fluorescence emission and CD spectra similar to L1 overexpressed in rich media. On the other hand, the mmL1\u2212Zn sample did not bind to a Q\u2010Sepharose column, and gel filtration studies demonstrated that this protein was monomeric. The mmL1\u2212Zn sample exhibited a lower <jats:italic>k<\/jats:italic><jats:sub>cat<\/jats:sub> value, bound less Zn(II), and yielded fluorescence emission and CD spectra consistent with this enzyme being folded improperly. Taken together, these data demonstrate that the proper folding of L1 requires the presence of Zn(II) and suggest that in vitro, thermodynamic metal binding studies do not accurately reflect physiological metal incorporation into L1.<\/jats:p>","DOI":"10.1110\/ps.04742704","type":"journal-article","created":{"date-parts":[[2004,7,7]],"date-time":"2004-07-07T01:14:27Z","timestamp":1089162867000},"page":"2236-2243","source":"Crossref","is-referenced-by-count":16,"title":["In vivo folding of recombinant metallo\u2010\u03b2\u2010lactamase L1 requires the presence of Zn(II)"],"prefix":"10.1002","volume":"13","author":[{"given":"Gopalraj","family":"Periyannan","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrick J.","family":"Shaw","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tara","family":"Sigdel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael W.","family":"Crowder","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2009,1]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1098\/rstb.1980.0049"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0168-6445(03)00041-X"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.33.3.259"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.33.3.264"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.33.3.271"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1086\/514922"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.39.6.1211"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2091-3-4"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1367-5931(99)00070-8"},{"key":"e_1_2_7_11_1","doi-asserted-by":"crossref","first-page":"915","DOI":"10.2174\/1381612805666230112193307","article-title":"Class B \u03b2\u2010lactamases: The importance of being metallic","volume":"5","author":"Cricco J.A.","year":"1999","journal-title":"Curr. 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