{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T14:01:52Z","timestamp":1789999312637,"version":"4.0.1"},"reference-count":0,"publisher":"Portland Press Ltd.","issue":"2","content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[1993,6,1]]},"abstract":"<jats:p>The cDNA of the human GM2-activator protein was cloned into the expression vector pHX17. The plasmid encodes a fusion protein with a hexahistidine tail and a Factor Xa cleavage site at its N-terminus. The recombinant protein was purified from cell homogenates under denaturing conditions by metal-ion affinity chromatography in a single step and then was refolded. The hexahistidine tail could be removed when desired by digestion with Factor Xa. In a functional assay, the GM2-activator thus generated from Escherichia coli and renatured, with or without the hexahistidine tail, was as active as the native GM2-activator protein that was purified from human tissue. When added to the culture medium, the recombinant carbohydrate-free GM2-activator, carrying the hexahistidine tail, could be taken up efficiently and restored the degradation of ganglioside GM2 to normal rates in mutant fibroblasts with the AB variant of GM2-gangliosidosis, which is characterized by a genetic defect in the GM2-activator protein. The prokaryotic expression system is useful for producing milligram quantities of a pure and functionally active GM2-activator.<\/jats:p>","DOI":"10.1042\/bj2920571","type":"journal-article","created":{"date-parts":[[2015,8,10]],"date-time":"2015-08-10T21:32:45Z","timestamp":1439242365000},"page":"571-576","update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":56,"title":["Over-expression of a functionally active human GM2-activator protein in <i>Escherichia coli<\/i>"],"prefix":"10.1042","volume":"292","author":[{"given":"H","family":"Klima","sequence":"first","affiliation":[{"name":"Institut f\u00fcr Organische Chemie und Biochemie, Gerhard-Domagk-Stra\u00dfe 1, D-5300 Bonn 1, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A","family":"Klein","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Organische Chemie und Biochemie, Gerhard-Domagk-Stra\u00dfe 1, D-5300 Bonn 1, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G","family":"van Echten","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Organische Chemie und Biochemie, Gerhard-Domagk-Stra\u00dfe 1, D-5300 Bonn 1, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G","family":"Schwarzmann","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Organische Chemie und Biochemie, Gerhard-Domagk-Stra\u00dfe 1, D-5300 Bonn 1, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"K","family":"Suzuki","sequence":"additional","affiliation":[{"name":"Brain and Development Research Center, University of North Carolina School of Medicine, Chapel Hill, NC 27599, U.S.A."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"K","family":"Sandhoff","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Organische Chemie und Biochemie, Gerhard-Domagk-Stra\u00dfe 1, D-5300 Bonn 1, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"288","container-title":["Biochemical Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/portlandpress.com\/biochemj\/article-pdf\/292\/2\/571\/610759\/bj2920571.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/portlandpress.com\/biochemj\/article-pdf\/292\/2\/571\/610759\/bj2920571.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,11,23]],"date-time":"2021-11-23T21:03:15Z","timestamp":1637701395000},"score":1,"resource":{"primary":{"URL":"https:\/\/portlandpress.com\/biochemj\/article\/292\/2\/571\/30250\/Over-expression-of-a-functionally-active-human-GM2"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1993,6,1]]},"references-count":0,"journal-issue":{"issue":"2","published-print":{"date-parts":[[1993,6,1]]}},"URL":"https:\/\/doi.org\/10.1042\/bj2920571","relation":{},"ISSN":["0264-6021","1470-8728"],"issn-type":[{"value":"0264-6021","type":"print"},{"value":"1470-8728","type":"electronic"}],"subject":[],"published":{"date-parts":[[1993,6,1]]}}}