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Conformational changes in formylglycine-generating enzyme during the catalytic cycle: Role of reducing agent and calcium

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Conformational changes in formylglycine-generating enzyme during the catalytic cycle: Role of reducing agent and calcium

Md Sarfaraz Alam (Autor)

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ISBN-13 (Printausgabe) 9783736991101
ISBN-13 (E-Book) 9783736981102
Sprache Englisch
Seitenanzahl 158
Umschlagkaschierung glänzend
Auflage 1. Aufl.
Erscheinungsort Göttingen
Promotionsort Göttingen
Erscheinungsdatum 21.09.2015
Allgemeine Einordnung Dissertation
Fachbereiche Biochemie, Molekularbiologie, Gentechnologie
Schlagwörter Formylglycine generation enzyme, Multiple sulfatase deficiency, in vitro activity
Beschreibung

Formylglycine generation, a unique process necessary for the activation of sulfatases, is a cotranslational event conserved from pro- to eukaryotes and is defined by the modification of the cysteine residue in the CxPxR motif of largely unfolded sulfatase polypeptides to Cα-formylglycine (FGly). This process is confined to the endoplasmic reticulum (ER) in eukaryotes and catalyzed by the formylglycine-generating enzyme (FGE), a resident protein of the ER. Mutations in FGE, that impair FGly generation, lead to production of inactive sulfatases that manifests into an inherited metabolic disorder termed multiple sulfatase deficiency (MSD) in humans. Although FGE has been proposed to function as a cofactor independent monooxygenase, the precise mechanism is not yet clear. In the present study, a baculovirus-based method for expression and purification of recombinant FGE from cultured insect cells was established and the recombinant FGE was characterized in more detail.