{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T21:10:00Z","timestamp":1791234600935,"version":"4.1.0"},"reference-count":47,"publisher":"Portland Press Ltd.","issue":"3","content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2010,2,1]]},"abstract":"<jats:p>A putative GDP-Man PP (guanidine diphosphomannose pyrophosphorylase) gene from Trypanosoma brucei (TbGDP-Man PP) was identified in the genome and subsequently cloned, sequenced and recombinantly expressed, and shown to be a catalytically active dimer. Kinetic analysis revealed a Vmax of 0.34 \u03bcmol\/min per mg of protein and Km values of 67 \u03bcM and 12 \u03bcM for GTP and mannose 1-phosphate respectively. Further kinetic studies showed GDP-Man was a potent product feedback inhibitor. RNAi (RNA interference) of the cytosolic TbGDP-Man PP showed that mRNA levels were reduced to ~20% of wild-type levels, causing the cells to die after 3\u20134 days, demonstrating that TbGDP-Man PP is essential in the bloodstream form of T. brucei and thus a potential drug target. The RNAi-induced parasites have a greatly reduced capability to form GDP-Man, leading ultimately to a reduction in their ability to synthesize their essential GPI (glycosylphosphatidylinositol) anchors. The RNAi-induced parasites also showed aberrant N-glycosylation of their major cell-surface glycoprotein, variant surface glycoprotein, with loss of the high-mannose Man9GlcNAc2 N-glycosylation at Asn428 and formation of complex N-glycans at Asn263.<\/jats:p>","DOI":"10.1042\/bj20090896","type":"journal-article","created":{"date-parts":[[2009,11,18]],"date-time":"2009-11-18T18:35:18Z","timestamp":1258569318000},"page":"603-614","update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":26,"title":["GDP-mannose pyrophosphorylase is essential in the bloodstream form of <i>Trypanosoma brucei<\/i>"],"prefix":"10.1042","volume":"425","author":[{"given":"Helen","family":"Denton","sequence":"first","affiliation":[{"name":"Biomolecular Sciences Research Complex, The North Haugh, The University, St Andrews, Fife KY16 9ST, Scotland, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stewart","family":"Fyffe","sequence":"additional","affiliation":[{"name":"Biomolecular Sciences Research Complex, The North Haugh, The University, St Andrews, Fife KY16 9ST, Scotland, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Terry\u00a0K.","family":"Smith","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"288","published-online":{"date-parts":[[2010,1,15]]},"reference":[{"key":"2021112214183789300_B1","first-page":"57","article-title":"Exploring the biological role of glycans","volume-title":"Essentials of Glycobiology","author":"Varki","year":"1999"},{"key":"2021112214183789300_B2","doi-asserted-by":"crossref","first-page":"6866","DOI":"10.1046\/j.1432-1033.2000.01781.x","article-title":"Cloning, expression and characterization of the pig liver GDP-mannose pyrophosphorylase. 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