{"id":8892,"date":"2022-06-01T06:00:39","date_gmt":"2022-05-31T21:00:39","guid":{"rendered":"https:\/\/www.emukk.com\/WP\/?p=8892"},"modified":"2022-05-29T07:51:18","modified_gmt":"2022-05-28T22:51:18","slug":"a-novel-method-to-detect-o-glcnacylated-proteins-with-a-disaccharide-tag-and-wja-lectin","status":"publish","type":"post","link":"https:\/\/www.emukk.com\/WP\/en\/a-novel-method-to-detect-o-glcnacylated-proteins-with-a-disaccharide-tag-and-wja-lectin\/","title":{"rendered":"A novel method to detect O-GlcNAcylated proteins with a disaccharide-tag and WJA lectin"},"content":{"rendered":"<p>A group from Faculty of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwa, Chiba, Japan, etc. has reported a novel method to detect O-GlcNAcylated proteins specifically and with high sensitivity.<br \/>\n<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC9126400\/\">https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC9126400\/<\/a><\/p>\n<p>O-GlcNAc modification of serine or threonine occurs only on nuclear or cytoplasmic proteins. This modification is significantly different from N-glycosylation or O-glycosylation from several perspectives.<br \/>\n(1) O-GlcNAcylation occurs in the nucleus or cytoplasm of the cell, whereas general glycosylation occurs in the luminal regions of the endoplasmic reticulum and Golgi apparatus.<br \/>\n(2) O-GlcNAcylation is a reversible reaction via the actions of O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA), although N- and O-glycosylation are irreversible.<br \/>\n(3) general N- and O-glycans on proteins are sequentially processed and elongated with many types of glycosyltransferases. However, O-GlcNAc residues cannot be modified by other monosaccharides.<\/p>\n<p>In this study, to detect O-GlcNAc modification of proteins, especially in the nucleus of the cell, expression vectors encoding soluble \u03b24GalNAc-TA were constructed with a nuclear localization signal (NLS) from the SV40 large T antigen at the N- or C-terminus, respectively, and let \u03b24GalNAc transferase transfer GalNAc to the GlcNAc residue of the O-GlcNAcylated proteins forming GalNAc\u03b21-4GlcNAc disaccharide. Then, Wisteria japonica agglutinin (WJA) was used as a probe to detect GalNAc\u03b21-4GlcNAc disaccharide, because WJA is highly specific for GalNAc\u03b21-4GlcNAc.<br \/>\nThis method was named as <strong>disaccharide-tag method<\/strong>.<\/p>\n<p><a href=\"https:\/\/www.emukk.com\/WP\/wp-content\/uploads\/2022\/06\/disaccharide-tag-method.png\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone size-medium wp-image-8889\" src=\"https:\/\/www.emukk.com\/WP\/wp-content\/uploads\/2022\/06\/disaccharide-tag-method-300x138.png\" alt=\"\" width=\"300\" height=\"138\" srcset=\"https:\/\/www.emukk.com\/WP\/wp-content\/uploads\/2022\/06\/disaccharide-tag-method-300x138.png 300w, https:\/\/www.emukk.com\/WP\/wp-content\/uploads\/2022\/06\/disaccharide-tag-method-600x276.png 600w, https:\/\/www.emukk.com\/WP\/wp-content\/uploads\/2022\/06\/disaccharide-tag-method.png 733w\" sizes=\"auto, (max-width: 300px) 100vw, 300px\" \/><\/a><\/p>\n<p>The binding between GalNAc-GlcNAc used in this method and WJA lectin has a stronger Ka of 1.4 x 10<sup>5<\/sup> M<sup>-1<\/sup> which enables higher sensitivity than any other methods. In addition, it should be emphasized that sugar chains with the GalNAc-GlcNAc sequence were hardly expressed in animal cells. The GalNAc-GlcNAc structure is found at the non-reducing end of the N-type sugar chain of pituitary glycoprotein hormones (leutropin, thyrotropin, follitropin), which serves as a clearance signal from the blood stream. These glycoprotein hormones are present only in the pituitary gland and are not expressed in other tissues and cells.<\/p>\n","protected":false},"excerpt":{"rendered":"<p>A group from Faculty of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo,<\/p><\/div>\n<div class=\"blog-btn\"><a href=\"https:\/\/www.emukk.com\/WP\/en\/a-novel-method-to-detect-o-glcnacylated-proteins-with-a-disaccharide-tag-and-wja-lectin\/\" class=\"home-blog-btn\">\u7d9a\u304d\u3092\u8aad\u3080<\/a><\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[7],"tags":[],"class_list":["post-8892","post","type-post","status-publish","format-standard","hentry","category-technology-en"],"aioseo_notices":[],"_links":{"self":[{"href":"https:\/\/www.emukk.com\/WP\/wp-json\/wp\/v2\/posts\/8892","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.emukk.com\/WP\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.emukk.com\/WP\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.emukk.com\/WP\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.emukk.com\/WP\/wp-json\/wp\/v2\/comments?post=8892"}],"version-history":[{"count":1,"href":"https:\/\/www.emukk.com\/WP\/wp-json\/wp\/v2\/posts\/8892\/revisions"}],"predecessor-version":[{"id":8893,"href":"https:\/\/www.emukk.com\/WP\/wp-json\/wp\/v2\/posts\/8892\/revisions\/8893"}],"wp:attachment":[{"href":"https:\/\/www.emukk.com\/WP\/wp-json\/wp\/v2\/media?parent=8892"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.emukk.com\/WP\/wp-json\/wp\/v2\/categories?post=8892"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.emukk.com\/WP\/wp-json\/wp\/v2\/tags?post=8892"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}