{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T13:41:18Z","timestamp":1791380478231,"version":"4.3.3"},"reference-count":34,"publisher":"Portland Press Ltd.","issue":"2","content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2006,3,1]]},"abstract":"<jats:p>Failure to detoxify the intermediary metabolite glyoxylate in human hepatocytes underlies the metabolic pathology of two potentially lethal hereditary calcium oxalate kidney stone diseases, PH (primary hyperoxaluria) types\u00a01 and 2. In order to define more clearly the roles of enzymes involved in the metabolism of glyoxylate, we have established singly, doubly and triply transformed CHO (Chinese-hamster ovary) cell lines, expressing all combinations of normal human AGT (alanine:glyoxylate aminotransferase; the enzyme deficient in PH1), GR\/HPR (glyoxylate\/hydroxypyruvate reductase; the enzyme deficient in PH2), and GO (glycolate oxidase). We have embarked on the preliminary metabolic analysis of these transformants by studying the indirect toxicity of glycolate as a simple measure of the net intracellular production of glyoxylate. Our results show that glycolate is toxic only to those cells expressing GO and that this toxicity is diminished when AGT and\/or GR\/HPR are expressed in addition to GO. This finding indicates that we have been able to reconstruct the glycolate\u2192glyoxylate, glyoxylate\u2192glycine, and glyoxylate\u2192glycolate metabolic pathways, catalysed by GO, AGT, and GR\/HPR respectively, in cells that do not normally express them. These results are compatible with the findings in PH1 and PH2, in which AGT and GR\/HPR deficiencies lead to increased oxalate synthesis, due to the failure to detoxify its immediate precursor glyoxylate. These CHO cell transformants have a potential use as a cell-based bioassay for screening small molecules that stabilize AGT or GR\/HPR and might have use in the treatment of PH1 or PH2.<\/jats:p>","DOI":"10.1042\/bj20051397","type":"journal-article","created":{"date-parts":[[2006,3,2]],"date-time":"2006-03-02T09:09:03Z","timestamp":1141290543000},"page":"409-416","update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":65,"title":["Reconstruction of human hepatocyte glyoxylate metabolic pathways in stably transformed Chinese-hamster ovary cells"],"prefix":"10.1042","volume":"394","author":[{"given":"Joseph\u00a0T.","family":"Behnam","sequence":"first","affiliation":[{"name":"Department of Biology, University College London, London WC1E 6BT, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emma\u00a0L.","family":"Williams","sequence":"additional","affiliation":[{"name":"Department of Clinical Biochemistry, University College London Hospitals, London W1T 4EU, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susanne","family":"Brink","sequence":"additional","affiliation":[{"name":"Department of Biology, University College London, London WC1E 6BT, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gill","family":"Rumsby","sequence":"additional","affiliation":[{"name":"Department of Clinical Biochemistry, University College London Hospitals, London W1T 4EU, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christopher\u00a0J.","family":"Danpure","sequence":"additional","affiliation":[{"name":"Department of Biology, University College London, London WC1E 6BT, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"288","published-online":{"date-parts":[[2006,2,10]]},"reference":[{"key":"2021112213552044000_B1","first-page":"3323","article-title":"Primary hyperoxaluria","volume-title":"The Metabolic and Molecular Bases of Inherited Disease","author":"Danpure","year":"2001"},{"key":"2021112213552044000_B2","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1111\/j.1432-1033.1976.tb10219.x","article-title":"L-\u03b1-hydroxyacid oxidase isozymes. Purification and properties","volume":"63","author":"Duley","year":"1976","journal-title":"Eur. J. Biochem."},{"key":"2021112213552044000_B3","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1007\/BF00982670","article-title":"Electron microscopic cytochemical localization of \u03b1-hydroxyacid oxidase in rat liver. Association with the crystalline core and matrix of peroxisomes","volume":"85","author":"Angermuller","year":"1986","journal-title":"Histochemistry"},{"key":"2021112213552044000_B4","first-page":"43","article-title":"Enzymology of endogenous oxalate formation","volume":"5","author":"Bais","year":"1987","journal-title":"Adv. Clin. Enzymol."},{"key":"2021112213552044000_B5","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/0006-2944(80)90051-4","article-title":"The organ distribution of human alanine-2-oxoglutarate aminotransferase and alanine-glyoxylate aminotransferase","volume":"23","author":"Kamoda","year":"1980","journal-title":"Biochem. Med."},{"key":"2021112213552044000_B6","doi-asserted-by":"crossref","first-page":"1285","DOI":"10.1177\/36.10.3418107","article-title":"Immunocytochemical localization of human hepatic alanine: glyoxylate aminotransferase in control subjects and patients with primary hyperoxaluria type\u00a01","volume":"36","author":"Cooper","year":"1988","journal-title":"J. Histochem. Cytochem."},{"key":"2021112213552044000_B7","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1177\/000456329803500114","article-title":"Kinetic analysis and tissue distribution of human D-glycerate dehydrogenase\/glyoxylare reductase and its relevance to the diagnosis of primary hyperoxaluria type\u00a02","volume":"35","author":"Giafi","year":"1998","journal-title":"Ann. Clin. Biochem."},{"key":"2021112213552044000_B8","doi-asserted-by":"crossref","first-page":"497","DOI":"10.1002\/humu.9200","article-title":"Molecular analysis of the glyoxylate reductase (GRHPR) gene and description of mutations underlying primary hyperoxaluria type\u00a02","volume":"22","author":"Cregeen","year":"2003","journal-title":"Hum. Mutat."},{"key":"2021112213552044000_B9","doi-asserted-by":"crossref","first-page":"626","DOI":"10.1042\/bst0160626","article-title":"Hepatic D-glycerate dehydrogenase and glyoxylate reductase deficiency in primary hyperoxaluria type\u00a02","volume":"16","author":"Mistry","year":"1988","journal-title":"Biochem. Soc. Trans."},{"key":"2021112213552044000_B10","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1016\/0014-5793(86)80563-4","article-title":"Peroxisomal alanine:glyoxylate aminotransferase deficiency in primary hyperoxaluria type\u00a0I","volume":"201","author":"Danpure","year":"1986","journal-title":"FEBS Lett."},{"key":"2021112213552044000_B11","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1056\/NEJM196802012780502","article-title":"L-glyceric aciduria. A new genetic variant of primary hyperoxaluria","volume":"278","author":"Williams","year":"1968","journal-title":"N. Engl. J. Med."},{"key":"2021112213552044000_B12","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1016\/0304-4165(83)90146-0","article-title":"Oxalate synthesis from (14C1)glycollate and (14C1)glyoxylate in the hepatectomized rat","volume":"757","author":"Farinelli","year":"1983","journal-title":"Biochim. Biophys. Acta"},{"key":"2021112213552044000_B13","doi-asserted-by":"crossref","first-page":"438","DOI":"10.1016\/0003-9861(72)90361-X","article-title":"The metabolism of oxalate precursors in isolated perfused rat livers","volume":"153","author":"Liao","year":"1972","journal-title":"Arch. Biochem. Biophys."},{"key":"2021112213552044000_B14","first-page":"29","article-title":"The metabolic production of oxalate from xylitol: activities of transketolase, transaldolase, fructokinase and aldolase in liver, kidney, brain, heart and muscle in the rat, mouse, guinea pig, rabbit and human","volume":"28","author":"James","year":"1985","journal-title":"Int. J. Vitam. Nutr. Res. Suppl."},{"key":"2021112213552044000_B15","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1042\/bj3520409","article-title":"The peroxisomal targeting sequence type\u00a01 receptor, Pex5p, and the peroxisomal import efficiency of alanine:glyoxylate aminotransferase","volume":"352","author":"Knott","year":"2000","journal-title":"Biochem. J."},{"key":"2021112213552044000_B16","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1016\/S0167-4781(99)00105-0","article-title":"Identification and expression of a cDNA for human hydroxypyruvate\/glyoxylate reductase","volume":"1446","author":"Rumsby","year":"1999","journal-title":"Biochim. Biophys. Acta"},{"key":"2021112213552044000_B17","doi-asserted-by":"crossref","first-page":"246","DOI":"10.1016\/S0167-4781(00)00161-5","article-title":"Identification and expression of a cDNA for human glycolate oxidase","volume":"1493","author":"Williams","year":"2000","journal-title":"Biochim. Biophys. Acta"},{"key":"2021112213552044000_B18","doi-asserted-by":"crossref","first-page":"36415","DOI":"10.1074\/jbc.M006693200","article-title":"Functional synergism between the most common polymorphism in human alanine:glyoxylate aminotransferase and four of the most common disease-causing mutations","volume":"275","author":"Lumb","year":"2000","journal-title":"J. Biol. Chem."},{"key":"2021112213552044000_B19","doi-asserted-by":"crossref","first-page":"400","DOI":"10.1177\/000456329703400411","article-title":"A semiautomated alanine:glyoxylate aminotransferase assay for the tissue diagnosis of primary hyperoxaluria type\u00a01","volume":"34","author":"Rumsby","year":"1997","journal-title":"Ann. Clin. Biochem."},{"key":"2021112213552044000_B20","unstructured":"Williams\n              E.\u00a0L.\n            \n          \n          Human liver glycolate oxidase \u2013 gene identification and protein studies\n          2003\n          University of London\n          PhD Thesis"},{"key":"2021112213552044000_B21","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1083\/jcb.131.1.95","article-title":"Mammalian alanine:glyoxylate aminotransferase 1 is imported into peroxisomes via the PTS1 translocation pathway. Increased degeneracy and context specificity of the mammalian PTS1 motif and implications for the peroxisome-to-mitochondrion mistargeting of AGT in primary hyperoxaluria type\u00a01","volume":"131","author":"Motley","year":"1995","journal-title":"J. Cell Biol."},{"key":"2021112213552044000_B22","doi-asserted-by":"crossref","first-page":"12590","DOI":"10.1074\/jbc.275.17.12590","article-title":"Identification and characterization of HAOX1, HAOX2, and HAOX3, three human peroxisomal 2-hydroxy acid oxidases","volume":"275","author":"Jones","year":"2000","journal-title":"J. Biol. Chem."},{"key":"2021112213552044000_B23","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1053\/ajkd.2001.26099","article-title":"Ethylene glycol-mediated tubular injury: identification of critical metabolites and injury pathways","volume":"38","author":"Poldelski","year":"2001","journal-title":"Am. J. Kidney Dis."},{"key":"2021112213552044000_B24","doi-asserted-by":"crossref","first-page":"517","DOI":"10.1042\/bj2680517","article-title":"Human peroxisomal L-alanine: glyoxylate aminotransferase. Evolutionary loss of a mitochondrial targeting signal by point mutation of the initiation codon","volume":"268","author":"Takada","year":"1990","journal-title":"Biochem. J."},{"key":"2021112213552044000_B25","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1007\/BF01802035","article-title":"Enzymological characterization of a feline analogue of primary hyperoxaluria type\u00a02: a model for the human disease","volume":"12","author":"Danpure","year":"1989","journal-title":"J. Inherited Metab. Dis."},{"key":"2021112213552044000_B26","doi-asserted-by":"crossref","first-page":"1335","DOI":"10.1002\/j.1460-2075.1986.tb04364.x","article-title":"Mitochondrial targeting sequences may form amphiphilic helices","volume":"5","author":"von\u00a0Heijne","year":"1986","journal-title":"EMBO J."},{"key":"2021112213552044000_B27","doi-asserted-by":"crossref","first-page":"4115","DOI":"10.1002\/j.1460-2075.1993.tb06095.x","article-title":"Functional cooperation of mitochondrial protein import receptors in yeast","volume":"12","author":"Ramage","year":"1993","journal-title":"EMBO J."},{"key":"2021112213552044000_B28","doi-asserted-by":"crossref","first-page":"863","DOI":"10.1146\/annurev.biochem.66.1.863","article-title":"Protein import into mitochondria","volume":"66","author":"Neupert","year":"1997","journal-title":"Annu. Rev. Biochem."},{"key":"2021112213552044000_B29","doi-asserted-by":"crossref","first-page":"1129","DOI":"10.1073\/pnas.96.3.1129","article-title":"Role of mitochondrial lactate dehydrogenase and lactate oxidation in the intracellular lactate shuttle","volume":"96","author":"Brooks","year":"1999","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2021112213552044000_B30","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1016\/j.bbapap.2005.08.004","article-title":"A preliminary account of the properties of recombinant human glyoxylate reductase (GRHPR), LDHA and LDHB with glyoxylate, and their potential roles in its metabolism","volume":"1753","author":"Mdluli","year":"2005","journal-title":"Biochim. Biophys. Acta."},{"key":"2021112213552044000_B31","doi-asserted-by":"crossref","first-page":"2341","DOI":"10.1083\/jcb.111.6.2341","article-title":"Identification of mutations associated with peroxisome-to-mitochondrion mistargeting of alanine\/glyoxylate aminotransferase in primary hyperoxaluria type\u00a01","volume":"111","author":"Purdue","year":"1990","journal-title":"J. Cell Biol."},{"key":"2021112213552044000_B32","doi-asserted-by":"crossref","first-page":"1704","DOI":"10.1111\/j.1523-1755.2005.00267.x","article-title":"Pyridoxine effect in type\u00a01 primary hyperoxaluria is associated with the commonest mutant allele","volume":"67","author":"Monico","year":"2005","journal-title":"Kidney Int."},{"key":"2021112213552044000_B33","doi-asserted-by":"crossref","first-page":"1345","DOI":"10.1083\/jcb.108.4.1345","article-title":"An enzyme trafficking defect in two patients with primary hyperoxaluria type\u00a01: peroxisomal alanine\/glyoxylate aminotransferase rerouted to mitochondria","volume":"108","author":"Danpure","year":"1989","journal-title":"J. Cell Biol."},{"key":"2021112213552044000_B34","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1042\/bj20030371","article-title":"Correction of an enzyme trafficking defect in hereditary kidney stone disease in vitro","volume":"374","author":"Lumb","year":"2003","journal-title":"Biochem. J."}],"container-title":["Biochemical Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/portlandpress.com\/biochemj\/article-pdf\/394\/2\/409\/641558\/bj3940409.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/portlandpress.com\/biochemj\/article-pdf\/394\/2\/409\/641558\/bj3940409.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,11,22]],"date-time":"2021-11-22T15:53:32Z","timestamp":1637596412000},"score":1,"resource":{"primary":{"URL":"https:\/\/portlandpress.com\/biochemj\/article\/394\/2\/409\/43713\/Reconstruction-of-human-hepatocyte-glyoxylate"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2006,2,10]]},"references-count":34,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2006,3,1]]}},"URL":"https:\/\/doi.org\/10.1042\/bj20051397","relation":{},"ISSN":["0264-6021","1470-8728"],"issn-type":[{"value":"0264-6021","type":"print"},{"value":"1470-8728","type":"electronic"}],"subject":[],"published":{"date-parts":[[2006,2,10]]}}}