title: Overexpression of inactive arylsulphatase mutants and in vitro activation by light-dependent oxidation with vanadate.
abstract: Arylsulphatases B (ASB) and A (ASA) are subject to a unique post-translational modification that is required for their function. The modification reaction, conversion of an active-site cysteine into a formylglycine, becomes saturated when these enzymes are overexpressed. We have removed the possibility of in vivo modification by expressing mutants of ASB and ASA in which the active-site cysteine is substituted with a serine. These mutants are expressed much more efficiently when compared with the native enzymes under identical conditions. The purified ASB mutant can then be converted into catalytically active ASB in vitro using vanadate and light.
authors.affiliation: BioMarin Pharmaceutical Inc., 371 Bel Marin Keys Blvd., Novato, CA 94949, USA.
authors.forename: Terri M
authors.initials: TM
authors.lastname: Christianson
authors.forename: Chris M
authors.initials: CM
authors.lastname: Starr
authors.forename: Todd C
authors.initials: TC
authors.lastname: Zankel
mesh_terms: D000818:Animals; D001192:Arylsulfatases / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*; D016466:CHO Cells / Q000737:chemistry / Q000378:metabolism; D002460:Cell Line; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics; D004789:Enzyme Activation; D006801:Humans; D008027:Light; D008099:Liver / Q000201:enzymology; D020125:Mutation, Missense / Q000235:genetics; D010084:Oxidation-Reduction; D014162:Transfection / Q000379:methods; D014638:Vanadates / Q000737:chemistry / Q000378:metabolism*
chemical_list: D018076:DNA, Complementary; D014638:Vanadates; D001192:Arylsulfatases