{"id":118286,"date":"2021-01-10T19:23:15","date_gmt":"2021-01-11T03:23:15","guid":{"rendered":"https:\/\/lifeboat.com\/blog\/2021\/01\/engineered-sam-synthetases-for-enzymatic-generation-of-adomet-analogs-with-photocaging-groups-and-reversible-dna-modification-in-cascade-reactions"},"modified":"2021-01-10T19:23:15","modified_gmt":"2021-01-11T03:23:15","slug":"engineered-sam-synthetases-for-enzymatic-generation-of-adomet-analogs-with-photocaging-groups-and-reversible-dna-modification-in-cascade-reactions","status":"publish","type":"post","link":"https:\/\/lifeboat.com\/blog\/2021\/01\/engineered-sam-synthetases-for-enzymatic-generation-of-adomet-analogs-with-photocaging-groups-and-reversible-dna-modification-in-cascade-reactions","title":{"rendered":"Engineered SAM Synthetases for Enzymatic Generation of AdoMet Analogs with Photocaging Groups and Reversible DNA Modification in Cascade Reactions"},"content":{"rendered":"<p><a class=\"aligncenter blog-photo\" href=\"https:\/\/lifeboat.com\/blog.images\/engineered-sam-synthetases-for-enzymatic-generation-of-adomet-analogs-with-photocaging-groups-and-reversible-dna-modification-in-cascade-reactions3.jpg\"><\/a><\/p>\n<p>Methylation and demethylation of DNA, RNA and proteins has emerged as a major regulatory mechanism. Studying the function of these modifications would benefit from tools for their site\u2010specific inhibition and timed removal. <i>S <\/i>\u2010Adenosyl\u2010L\u2010methionine (AdoMet) analogs in combination with methyltransferases (MTases) have proven useful to map or block and release MTase target sites, however their enzymatic generation has been limited to aliphatic groups at the sulfur atom. We engineered a SAM synthetase from <i>Cryptosporidium hominis<\/i> (PC\u2010ChMAT) for efficient generation of AdoMet analogs with photocaging groups that are not accepted by any WT MAT reported to date. The crystal structure of PC\u2010ChMAT at 1.87 \u00c5 revealed how the photocaged AdoMet analog is accommodated and guided engineering of a thermostable MAT from <i>Methanocaldococcus jannaschii<\/i>. PC\u2010MATs were compatible with DNA\u2010 and RNA\u2010MTases, enabling sequence\u2010specific modification (\u201c<i>writing<\/i>\u201d) of plasmid DNA and light\u2010triggered removal (\u201c<i>erasing<\/i>\u201d).<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Methylation and demethylation of DNA, RNA and proteins has emerged as a major regulatory mechanism. Studying the function of these modifications would benefit from tools for their site\u2010specific inhibition and timed removal. S \u2010Adenosyl\u2010L\u2010methionine (AdoMet) analogs in combination with methyltransferases (MTases) have proven useful to map or block and release MTase target sites, however their [\u2026]<\/p>\n","protected":false},"author":427,"featured_media":0,"comment_status":"open","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[11,38],"tags":[],"class_list":["post-118286","post","type-post","status-publish","format-standard","hentry","category-biotech-medical","category-engineering"],"_links":{"self":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/posts\/118286","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/users\/427"}],"replies":[{"embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/comments?post=118286"}],"version-history":[{"count":0,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/posts\/118286\/revisions"}],"wp:attachment":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/media?parent=118286"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/categories?post=118286"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/tags?post=118286"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}