{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,6]],"date-time":"2026-03-06T14:02:00Z","timestamp":1772805720192,"version":"3.50.1"},"reference-count":33,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2008,12,31]],"date-time":"2008-12-31T00:00:00Z","timestamp":1230681600000},"content-version":"vor","delay-in-days":3836,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[1998,7]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Reverse transcriptase is an essential retroviral enzyme that uses RNA\u2010 and DNA\u2010directed DNA polymerase activities as well as an RNaseH activity to synthesize a double\u2010stranded DNA copy of the single\u2010stranded RNA genome. In an effort to obtain high\u2010resolution structural information regarding the polymerase active site of reverse transcriptase, we have pursued studies on a catalytic fragment from Moloney murine leukemia virus reverse transcriptase. DNA encoding the catalytic fragment, defined originally by limited proteolytic digestion, has been cloned, and the protein has been expressed and purified from <jats:italic>Escherichia coli<\/jats:italic>. The fragment obtained by limited proteolytic digestion and the bacterially expressed fragment retain polymerase activity. Crystallization studies involving nucleic acid complexes with a catalytic fragment from both sources are reported, including variables screened to improve crystals and cryocooling. Three crystal forms of catalytic fragment\u2010nucleic acid complexes have been characterized, which all contain at least two protein molecules in the asymmetric unit. As isolated, the catalytic fragment is monomeric. This analysis indicates that the enzyme dimerizes in the presence of nucleic acid.<\/jats:p>","DOI":"10.1002\/pro.5560070711","type":"journal-article","created":{"date-parts":[[2009,2,10]],"date-time":"2009-02-10T06:30:05Z","timestamp":1234247405000},"page":"1575-1582","source":"Crossref","is-referenced-by-count":17,"title":["Cloning, expression, and purification of a catalytic fragment of moloney murine leukemia virus reverse transcriptase: Crystallization of nucleic acid complexes"],"prefix":"10.1002","volume":"7","author":[{"given":"Dunming","family":"Sun","sequence":"first","affiliation":[]},{"given":"Sven","family":"Jessen","sequence":"additional","affiliation":[]},{"given":"Chunhui","family":"Liu","sequence":"additional","affiliation":[]},{"given":"Xiuping","family":"Liu","sequence":"additional","affiliation":[]},{"given":"Shabir","family":"Najmudin","sequence":"additional","affiliation":[]},{"given":"Millie M.","family":"Georgiadis","sequence":"additional","affiliation":[]}],"member":"311","published-online":{"date-parts":[[2008,12,31]]},"reference":[{"key":"e_1_2_1_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1046-2023(05)80150-1"},{"key":"e_1_2_1_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/elps.1150080203"},{"key":"e_1_2_1_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(19)86452-9"},{"key":"e_1_2_1_4_2","first-page":"760","article-title":"Collaborative computational project, number 4. 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The CCP4 suite: Programs for protein crystallography","volume":"50","author":"CCP4","year":"1994","journal-title":"Acta Crystallogr"},{"key":"e_1_2_1_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/nsb0595-407"},{"key":"e_1_2_1_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0969-2126(01)00168-X"},{"key":"e_1_2_1_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/nsb0495-303"},{"key":"e_1_2_1_8_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.89.2.748"},{"key":"e_1_2_1_9_1","first-page":"379","article-title":"Mechanistic implications from the structure of a catalytic fragment of Moloney murine leukemia virus reverse transcriptase","volume":"32","author":"Georgiadis MM","year":"1995","journal-title":"Structure"},{"key":"e_1_2_1_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(79)90357-X"},{"key":"e_1_2_1_11_1","first-page":"317","article-title":"Review: Retroviral reverse tranwriptase: Synthesis, structure, and function","volume":"32","author":"Goff SP","year":"1990","journal-title":"J Acquir lmmun Defic Syndrome"},{"key":"e_1_2_1_12_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi00015a013"},{"key":"e_1_2_1_13_1","first-page":"353","article-title":"Structure of unliganded HIV\u2010I reverse transcriptase at 2.7\u00c5, resolution: Implications of conformational changes for polymerization and inhibition mechanisms","volume":"4","author":"Hsiou Y","year":"1996","journal-title":"Strucrure"},{"key":"e_1_2_1_14_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.90.13.6320"},{"key":"e_1_2_1_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0969-2126(94)00087-5"},{"key":"e_1_2_1_16_1","doi-asserted-by":"publisher","DOI":"10.1107\/S0021889891004430"},{"key":"e_1_2_1_17_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1377403"},{"key":"e_1_2_1_17_2","first-page":"156","volume-title":"Reverse franscriptase","author":"LeGrice SFJ.","year":"1993"},{"key":"e_1_2_1_18_1","doi-asserted-by":"crossref","first-page":"727","DOI":"10.1128\/jvi.45.2.727-739.1983","article-title":"Mechanism of action of the endonuclease associated with the ah and bb forms of avian RNA tumor virus reverse transcriptase","volume":"45","author":"Leis H","year":"1983","journal-title":"J Virol"},{"key":"e_1_2_1_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/0022-2836(68)90205-2"},{"key":"e_1_2_1_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/0042-6822(74)90302-X"},{"key":"e_1_2_1_21_1","doi-asserted-by":"publisher","DOI":"10.1038\/nsb0396-267"},{"key":"e_1_2_1_22_1","doi-asserted-by":"publisher","DOI":"10.1038\/313762a0"},{"key":"e_1_2_1_23_1","first-page":"56","volume-title":"Proceedings of the CCP4 study weekend:Data collection and processing","author":"Otwinowsky Z.","year":"1993"},{"key":"e_1_2_1_24_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.7516580"},{"key":"e_1_2_1_25_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.1.430"},{"key":"e_1_2_1_26_1","doi-asserted-by":"publisher","DOI":"10.1038\/nsb0495-293"},{"key":"e_1_2_1_27_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.92.4.1222"},{"key":"e_1_2_1_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(17)39369-9"},{"key":"e_1_2_1_29_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.7528445"},{"key":"e_1_2_1_30_1","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.66.2.615-622.1992"},{"key":"e_1_2_1_31_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.90.4.1276"},{"key":"e_1_2_1_32_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0969-2126(94)00097-2"}],"container-title":["Protein Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fpro.5560070711","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/pro.5560070711","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,29]],"date-time":"2023-10-29T00:24:22Z","timestamp":1698539062000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/pro.5560070711"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1998,7]]},"references-count":33,"journal-issue":{"issue":"7","published-print":{"date-parts":[[1998,7]]}},"alternative-id":["10.1002\/pro.5560070711"],"URL":"https:\/\/doi.org\/10.1002\/pro.5560070711","archive":["Portico"],"relation":{},"ISSN":["0961-8368","1469-896X"],"issn-type":[{"value":"0961-8368","type":"print"},{"value":"1469-896X","type":"electronic"}],"subject":[],"published":{"date-parts":[[1998,7]]}}}