{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,1,12]],"date-time":"2025-01-12T05:10:55Z","timestamp":1736658655583,"version":"3.32.0"},"reference-count":0,"publisher":"International Union of Crystallography (IUCr)","issue":"4","license":[{"start":{"date-parts":[[1999,4,1]],"date-time":"1999-04-01T00:00:00Z","timestamp":922924800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/journals.iucr.org\/services\/copyrightpolicy.html"},{"start":{"date-parts":[[1999,4,1]],"date-time":"1999-04-01T00:00:00Z","timestamp":922924800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/journals.iucr.org\/services\/copyrightpolicy.html#TDM"}],"content-domain":{"domain":["iucr.org","wiley.com","iucrj.org"],"crossmark-restriction":true},"short-container-title":["Acta Crystallogr D Biol Crystallogr","Acta Cryst D","Acta Cryst Sect D","Acta Crystallogr D","Acta Crystallogr Sect D","Acta Crystallogr Sect D Biol Crystallogr","Acta Crystallogr D Biol Cryst"],"published-print":{"date-parts":[[1999,4,1]]},"abstract":"<jats:p>\u03b1-D-Glucuronidases cleave the \u03b1-1,2-glycosidic bond of the 4-<jats:italic>O<\/jats:italic>-methyl-\u03b1-D-glucuronic acid side chain in xylan. Of the xylan-debranching hydrolases, these enzymes are the least studied and characterized. The \u03b1-glucuronidase gene (<jats:italic>aguA<\/jats:italic>) from <jats:italic>Bacillus stearothermophilus<\/jats:italic> T-6 has been cloned, sequenced and overproduced in <jats:italic>Escherichia coli<\/jats:italic>. The gene encodes for a protein of 679 amino acids with a calculated molecular weight of 78480 and a p<jats:italic>I<\/jats:italic> of 5.42. \u03b1-Glucuronidase T-6 shows high homology to the \u03b1-glucuronidases of <jats:italic>Thermotoga maritima<\/jats:italic> (60% identity) and of <jats:italic>Trichoderma reesei<\/jats:italic> (44% identity). Based on the amino-acid sequence similarity, it is likely that these enzymes represent a new class of glycosyl hydrolases. Crystallographic studies of \u03b1-glucuronidase T-6 were initiated to study the mechanism of catalysis, as well as to provide a structural basis for rational introduction of enhanced thermostability by site-specific mutagenesis. In this report, the crystallization and preliminary crystallographic characterization of the native \u03b1-glucuronidase T-6 enzyme is described. Two crystal forms were found suitable for detailed crystal structure analysis. The T1 form was obtained by the vapour-diffusion method using PEG 4000 as a precipitant and 2-propanol as an organic additive. The crystals belong to a primitive tetragonal crystal system (space group <jats:italic>P<\/jats:italic>4<jats:sub>1<\/jats:sub>2<jats:sub>1<\/jats:sub>2 or <jats:italic>P<\/jats:italic>4<jats:sub>3<\/jats:sub>2<jats:sub>1<\/jats:sub>2) with unit-cell dimensions <jats:italic>a<\/jats:italic> = <jats:italic>b<\/jats:italic> = 76.1 and <jats:italic>c<\/jats:italic> = 331.2\u2005\u00c5. These crystals are mechanically strong, are stable in the X-ray beam and diffract X-rays to better than 2.4\u2005\u00c5 resolution. A full 3.0\u2005\u00c5 resolution diffraction data set (97.3% completeness, <jats:italic>R<\/jats:italic>\n                  <jats:sub>merge<\/jats:sub> 9.8%) has recently been collected on one crystal at room temperature using a rotating-anode X-ray source and an R-AXIS IIc imaging-plate detector. The M1 form was obtained and characterized by similar techniques. The best crystallization occurred at a slightly lower pH and a lower concentration of 2-propanol. The crystals belong to a primitive monoclinic crystal system (space group <jats:italic>P<\/jats:italic>2<jats:sub>1<\/jats:sub>) with unit-cell dimensions <jats:italic>a<\/jats:italic> = 65.8, <jats:italic>b<\/jats:italic> = 127.4, <jats:italic>c<\/jats:italic> = 96.6\u2005\u00c5 and \u03b2 = 97.9\u00b0. These crystals are also quite strong and stable, and diffract to better than 2.8\u2005\u00c5 resolution. A full 2.8\u2005\u00c5 resolution diffraction data set (96.2% completeness, <jats:italic>R<\/jats:italic>\n                  <jats:sub>merge<\/jats:sub> 7.6%) has recently been collected on one crystal at room temperature using the same R-AXIS IIc setup. Both forms are currently being used to obtain crystallographic phasing <jats:italic>via<\/jats:italic> isomorphous heavy-atom derivatives and selenomethionine MAD experiments.<\/jats:p>","DOI":"10.1107\/s0907444998012918","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T05:39:11Z","timestamp":1027748351000},"page":"869-872","update-policy":"https:\/\/doi.org\/10.1107\/cm_01","source":"Crossref","is-referenced-by-count":16,"title":["Crystallization and preliminary X-ray analysis of \u03b1-<scp>D<\/scp>-glucuronidase from <i>Bacillus stearothermophilus<\/i>\u00a0T-6"],"prefix":"10.1107","volume":"55","author":[{"given":"Anna","family":"Teplitsky","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Smadar","family":"Shulami","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sara","family":"Moryles","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Galia","family":"Zaide","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuval","family":"Shoham","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gil","family":"Shoham","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"329","published-online":{"date-parts":[[1999,4,1]]},"container-title":["Acta Crystallographica Section D Biological Crystallography"],"original-title":[],"link":[{"URL":"http:\/\/journals.iucr.org\/d\/issues\/1999\/04\/00\/gr0834\/gr0834.pdf","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"http:\/\/journals.iucr.org\/d\/issues\/1999\/04\/00\/gr0834\/gr0834.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,1,11]],"date-time":"2025-01-11T12:26:07Z","timestamp":1736598367000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.iucr.org\/paper?S0907444998012918"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1999,4,1]]},"references-count":0,"journal-issue":{"issue":"4","published-print":{"date-parts":[[1999,4]]}},"alternative-id":["S0907444998012918"],"URL":"https:\/\/doi.org\/10.1107\/s0907444998012918","relation":{},"ISSN":["0907-4449"],"issn-type":[{"type":"print","value":"0907-4449"}],"subject":[],"published":{"date-parts":[[1999,4,1]]},"assertion":[{"value":"Acta Crystallographica Section D: Biological Crystallography","order":0,"name":"journal","label":"Publication","explanation":{"URL":"http:\/\/journals.iucr.org\/d"},"group":{"name":"general","label":"General"}},{"value":"crystallization papers","order":1,"name":"content_type","label":"Content type","explanation":{"URL":"http:\/\/journals.iucr.org\/d\/services\/notesforauthors.html"},"group":{"name":"general","label":"General"}},{"value":"Available","URL":"http:\/\/scripts.iucr.org\/cgi-bin\/citedin?gr0834","order":2,"name":"article_metrics","label":"Article metrics","group":{"name":"general","label":"General"}},{"value":"Yes","order":0,"name":"peer_reviewed","label":"Peer reviewed","group":{"name":"peer_review","label":"Peer review"}},{"value":"Single blind","order":1,"name":"review_process","label":"Review process","group":{"name":"peer_review","label":"Peer review"}},{"value":"3 February 1998","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication history"}},{"value":"5 October 1998","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication history"}},{"value":"1 April 1999","order":2,"name":"published_online","label":"Published online","group":{"name":"publication_history","label":"Publication history"}},{"value":"\u00a9 1999 International Union of Crystallography","order":0,"name":"copyright","label":"Copyright","explanation":{"URL":"http:\/\/journals.iucr.org\/services\/copyrightpolicy.html"},"group":{"name":"copyright_and_licensing","label":"Copyright and licencing"}}]}}