{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,4]],"date-time":"2026-10-04T20:16:48Z","timestamp":1791145008966,"version":"4.1.0"},"reference-count":38,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2014,12,11]],"date-time":"2014-12-11T00:00:00Z","timestamp":1418256000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"NIH","award":["AI094335"],"award-info":[{"award-number":["AI094335"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2015,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Methylation of flavivirus RNA is vital for its stability and translation in the infected host cell. This methylation is mediated by the flavivirus methyltransferase (MTase), which methylates the N7 and 2\u2032\u2010O positions of the viral RNA cap by using S\u2010adenosyl\u2010<jats:sc>l<\/jats:sc>\u2010methionine (SAM) as a methyl donor. In this report, we demonstrate that SAM, in contrast to the reaction by\u2010product S\u2010adenosyl\u2010<jats:sc>l<\/jats:sc>\u2010homocysteine, which was assumed previously, is copurified with the Dengue (DNV) and West Nile virus MTases produced in <jats:italic>Escherichia coli<\/jats:italic> (<jats:italic>E. coli)<\/jats:italic>. This endogenous SAM can be removed by denaturation and refolding of the MTase protein. The refolded MTase of DNV serotype 3 (DNV3) displays methylation activity comparable to native enzyme, and its crystal structure at 2.1 \u00c5 is almost identical to that of native MTase. We characterized the binding of Sinefungin (SIN), a previously described SAM\u2010analog inhibitor of MTase function, to the native and refolded DNV3 MTase by isothermal titration calorimetry, and found that SIN binds to refolded MTase with more than 16 times the affinity of SIN binding to the MTase purified natively. Moreover, we show that SAM is also copurified with other flavivirus MTases, indicating that purification by refolding may be a generally applicable tool for studying flavivirus MTase inhibition.<\/jats:p>","DOI":"10.1002\/pro.2594","type":"journal-article","created":{"date-parts":[[2014,10,29]],"date-time":"2014-10-29T08:04:17Z","timestamp":1414569857000},"page":"117-128","source":"Crossref","is-referenced-by-count":21,"title":["Refolding of a fully functional flavivirus methyltransferase revealed that S\u2010adenosyl methionine but not S\u2010adenosyl homocysteine is copurified with flavivirus methyltransferase"],"prefix":"10.1002","volume":"24","author":[{"given":"Matthew B.","family":"Brecher","sequence":"first","affiliation":[{"name":"Wadsworth Center New York State Department of Health Albany New York 12208"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhong","family":"Li","sequence":"additional","affiliation":[{"name":"Wadsworth Center New York State Department of Health Albany New York 12208"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing","family":"Zhang","sequence":"additional","affiliation":[{"name":"Wadsworth Center New York State Department of Health Albany New York 12208"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hui","family":"Chen","sequence":"additional","affiliation":[{"name":"Wadsworth Center New York State Department of Health Albany New York 12208"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qishan","family":"Lin","sequence":"additional","affiliation":[{"name":"Center for Functional Genomics University at Albany Rensselaer New York 12144"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Binbin","family":"Liu","sequence":"additional","affiliation":[{"name":"Wadsworth Center New York State Department of Health Albany New York 12208"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongmin","family":"Li","sequence":"additional","affiliation":[{"name":"Wadsworth Center New York State Department of Health Albany New York 12208"},{"name":"Department of Biomedical Sciences School of Public Health, University at Albany, State University of New York Albany New York 12201\u20100509"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2014,12,11]]},"reference":[{"key":"e_1_2_7_2_1","unstructured":"Global Dengue(2014) Available at:http:\/\/www.cdc.gov\/dengue\/epidemiology\/index.html. 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