{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2023,10,17]],"date-time":"2023-10-17T05:47:07Z","timestamp":1697521627939},"reference-count":0,"publisher":"Portland Press Ltd.","issue":"3","content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[1997,2,1]]},"abstract":"<jats:p>Arg-136, Glu-137, Arg-138 and Arg-139 are conserved in all sequences of the 2-kinase domain of 6-phosphofructo-2-kinase\/fructose-2,6-bisphosphatase. Their role was studied by site-directed mutagenesis. All the mutations had little, if any, effect on fructose-2,6-bisphosphatase activity. Mutations of Arg-136 and Glu-137 into Ala caused only minor modifications of phosphofructo-2-kinase activity. In contrast, mutation of Arg-138 into Ala increased 280-fold the Km for fructose 6-phosphate of phosphofructo-2-kinase. Mutation of Arg-139 into Ala resulted in decreases in phosphofructo-2-kinase Vmax\/Km for MgATP and fructose 6-phosphate 600-fold and 5000-fold respectively. Mutation of Arg-139 into Lys and Gln increased the Km of phosphofructo-2-kinase for MgATP (20-fold and 25-fold respectively) and for fructose 6-phosphate (8-fold and 13-fold), and the IC50 for MgADP (30-fold and 50-fold) and for magnesium citrate (7-fold and 25-fold). However, these two mutations did not affect nucleotide binding, as measured by quenching of intrinsic fluorescence. The changes in kinetic properties induced by mutations could not be attributed to structural changes. It is proposed that Arg-138 is involved in fructose 6-phosphate binding and that Arg-139 is probably involved in the stabilization of the transition state and so participates in catalysis.<\/jats:p>","DOI":"10.1042\/bj3210609","type":"journal-article","created":{"date-parts":[[2015,8,10]],"date-time":"2015-08-10T21:52:36Z","timestamp":1439243556000},"page":"609-614","update-policy":"http:\/\/dx.doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Mutagenesis of charged residues in a conserved sequence in the 2-kinase domain of 6-phosphofructo-2-kinase\/fructose-2,6-bisphosphatase"],"prefix":"10.1042","volume":"321","author":[{"given":"Luc","family":"BERTRAND","sequence":"first","affiliation":[{"name":"Facult\u00e9s Universitaires Notre-Dame de la Paix, Department of Biology, rue de Bruxelles, 61, B-5000 Namur, Belgium"},{"name":"Hormone and Metabolic Research Unit, University of Louvain Medical School, and International Institute of Cellular and Molecular Pathology, Avenue Hippocrate, 75, B-1200 Brussels, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Didier","family":"VERTOMMEN","sequence":"additional","affiliation":[{"name":"Hormone and Metabolic Research Unit, University of Louvain Medical School, and International Institute of Cellular and Molecular Pathology, Avenue Hippocrate, 75, B-1200 Brussels, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ernest","family":"FEYTMANS","sequence":"additional","affiliation":[{"name":"Facult\u00e9s Universitaires Notre-Dame de la Paix, Department of Biology, rue de Bruxelles, 61, B-5000 Namur, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Attilio Di","family":"PIETRO","sequence":"additional","affiliation":[{"name":"Institut de Biologie et de Chimie des Prot\u00e9ines, Centre National de la Recherche Scientifique, Passage du Vercors, F-69367 Lyon cedex 07, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mark H.","family":"RIDER","sequence":"additional","affiliation":[{"name":"Hormone and Metabolic Research Unit, University of Louvain Medical School, and International Institute of Cellular and Molecular Pathology, Avenue Hippocrate, 75, B-1200 Brussels, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Louis","family":"HUE","sequence":"additional","affiliation":[{"name":"Hormone and Metabolic Research Unit, University of Louvain Medical School, and International Institute of Cellular and Molecular Pathology, Avenue Hippocrate, 75, B-1200 Brussels, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"288","container-title":["Biochemical Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/portlandpress.com\/biochemj\/article-pdf\/321\/3\/609\/623683\/bj3210609.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/portlandpress.com\/biochemj\/article-pdf\/321\/3\/609\/623683\/bj3210609.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,11,24]],"date-time":"2021-11-24T00:37:25Z","timestamp":1637714245000},"score":1,"resource":{"primary":{"URL":"https:\/\/portlandpress.com\/biochemj\/article\/321\/3\/609\/34207\/Mutagenesis-of-charged-residues-in-a-conserved"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1997,2,1]]},"references-count":0,"journal-issue":{"issue":"3","published-print":{"date-parts":[[1997,2,1]]}},"URL":"https:\/\/doi.org\/10.1042\/bj3210609","relation":{},"ISSN":["0264-6021","1470-8728"],"issn-type":[{"value":"0264-6021","type":"print"},{"value":"1470-8728","type":"electronic"}],"subject":[],"published":{"date-parts":[[1997,2,1]]}}}