Posttranslational generation of C-alpha-formylglycin in eukaryotic sulfatases: development of the biochemical approach for the characterisation and purification of the modifying enzymee

Borissenko, Ljudmila

dc.contributor.advisor	Figura, Kurt von Prof. Dr. Dr. h.c.	de
dc.contributor.author	Borissenko, Ljudmila	de
dc.date.accessioned	2012-05-16T12:12:03Z	de
dc.date.available	2013-01-30T23:50:30Z	de
dc.date.issued	2003-07-15	de
dc.identifier.uri	http://hdl.handle.net/11858/00-1735-0000-0006-B6BA-3	de
dc.identifier.uri	http://dx.doi.org/10.53846/goediss-1394
dc.format.mimetype	application/pdf	de
dc.language.iso	eng	de
dc.rights.uri	http://webdoc.sub.gwdg.de/diss/copyrdiss.htm	de
dc.title	Posttranslational generation of C-alpha-formylglycin in eukaryotic sulfatases: development of the biochemical approach for the characterisation and purification of the modifying enzymee	de
dc.type	doctoralThesis	de
dc.title.translated	Charakterisierung und Anreicherung der Enzyme, das Formylglycinreste in Sulfatasen bildet	de
dc.contributor.referee	Mösch, Hans-Ulrich Prof. Dr.	de
dc.date.examination	2003-01-30	de
dc.subject.dnb	500 Naturwissenschaften allgemein	de
dc.description.abstracteng	Sulfatases carry at their active site a C Ñ-formylglycine (FGly) residue that is essential for enzyme activity. The formyl group of this FGly is directly involved in sulfate ester cleavage. A defect in FGly formation in human is the cause of a recessively inherited disease called Multiple Sulfatase Deficiency, characterised by the synthesis of the catalytically inactive sulfatase polypeptides and by the accumulation of their unprocessed substrates in the lysosomes. FGly is generated by oxidation of a concerved cystein (pro- and eukaryotes) or serine residue (prokaryotes) comprised in the sequience motif C/S-x-P-x-R. In eukaryotes the modification is catalysed by lumenal components of the endoplasmic reticulum during or after protein translocation and prior to folding of sulfatases. Under in vitro conditions, using an enriched and soluble protein fraction, FGly formation could be observed under strictly posttranslational conditions and independent of a signal peptide. The modification reaction was characterised kinetically and with respect to cofactor requirement, albeit the acceptor of the reducing equivalents during cystein oxidation remains unknown.So far the enzymatic machinery involved in FGly modification could not be identified. We tried to purify it from the reticuloplasm of bovine pancreas microsomes using a number of chromatographic techniques with the following identification of proteins. We established several chromatographic protocols for separation of luminal proteins on different columns (ion exchanger, gel filtration, hydrophobic interaction, lectin chromatography, affinity chromatography etc.). FGly generating enzyme was characterised kinetically and biochemically. By combination of chromatographic protocols we purified the FGly generating activity and identified the bands visualised after separation of proteins by SDS PAGE. Several possible candidates on the role of FGly generating enzyme are identified.	de
dc.contributor.coReferee	Gradmann, Dietrich Prof. Dr.	de
dc.contributor.thirdReferee	Herrmann, Bernd Prof. Dr.	de
dc.subject.topic	Mathematics and Computer Science	de
dc.subject.ger	Sulfatasen	de
dc.subject.ger	Enzyme Modification	de
dc.subject.ger	Formylglycin	de
dc.subject.eng	sulfatase	de
dc.subject.eng	posttranslational modification	de
dc.subject.eng	formylglycin	de
dc.subject.bk	35.70 Biochemie	de
dc.subject.bk	42.13 Molekularbiologie	de
dc.identifier.urn	urn:nbn:de:gbv:7-webdoc-395-4	de
dc.identifier.purl	webdoc-395	de
dc.affiliation.institute	Medizinische Fakultät	de
dc.subject.gokfull	WA Biologie	de
dc.identifier.ppn	374622795	de

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