Schmidt, Bernhard

[["dc.bibliographiccitation.firstpage","710"],["dc.bibliographiccitation.issue","4"],["dc.bibliographiccitation.journal","Biochimica et Biophysica Acta (BBA) - Molecular Cell Research"],["dc.bibliographiccitation.lastpage","725"],["dc.bibliographiccitation.volume","1793"],["dc.contributor.author","Dierks, Thomas"],["dc.contributor.author","Schlotawa, Lars"],["dc.contributor.author","Frese, Marc-Andre"],["dc.contributor.author","Radhakrishnan, Karthikeyan"],["dc.contributor.author","von Figura, Kurt"],["dc.contributor.author","Schmidt, Bernhard"],["dc.date.accessioned","2018-11-07T08:30:52Z"],["dc.date.available","2018-11-07T08:30:52Z"],["dc.date.issued","2009"],["dc.description.abstract","Multiple sulfatase deficiency (MSD), mucolipidosis (MIL) II/III and Niemann-Pick type C1 (NPC1) disease are rare but fatal lysosomal storage disorders caused by the genetic defect of non-lysosomal proteins. The NPC1 protein mainly localizes to late endosomes and is essential for cholesterol redistribution from endocytosed LDL to cellular membranes. NPC1 deficiency leads to lysosomal accumulation of a broad range of lipids. The precise functional mechanism of this membrane protein, however, remains puzzling. ML II, also termed I cell disease. and the less severe ML III result from deficiencies of the Golgi enzyme N-acetylglucosamine 1-phosphotransferase leading to a global defect of lysosome biogenesis. In patient cells, newly synthesized lysosomal proteins are not equipped with the critical lysosomal trafficking marker mannose 6-phosphate, thus escaping from lysosomal sorting at the trans Golgi network. MSD affects the entire sulfatase family, at least seven members of which are lysosomal enzymes that are specifically involved in the degradation of sulfated glycosaminoglycans, sulfolipids or other sulfated molecules. The combined deficiencies of all sulfatases result from a defective post-translational modification by the ER-localized formylglycine-generating enzyme (FGE), which oxidizes a specific cysteine residue to formylglycine, the catalytic residue enabling a unique mechanism of sulfate ester hydrolysis. This review gives an update on the molecular bases of these enigmatic diseases, which have been challenging researchers since many decades and so far led to a number of surprising findings that give deeper insight into both the cell biology and the pathobiochemistry underlying these complex disorders. In case of MSD, considerable progress has been made in recent years towards an understanding of disease-causing FGE mutations. First approaches to link molecular parameters with clinical manifestation have been described and even therapeutical options have been addressed. Further. the discovery of FGE as an essential sulfatase activating enzyme has considerable impact on enzyme replacement or gene therapy of lysosomal storage disorders caused by single sulfatase deficiencies. (C) 2008 Elsevier B.V. All rights reserved."],["dc.identifier.doi","10.1016/j.bbamcr.2008.11.015"],["dc.identifier.isi","000265369800011"],["dc.identifier.pmid","19124046"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/16996"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Elsevier Science Bv"],["dc.relation.issn","0167-4889"],["dc.title","Molecular basis of multiple sulfatase deficiency, mucolipidosis II/III and Niemann-Pick C1 disease - Lysosomal storage disorders caused by defects of non-lysosomal proteins"],["dc.type","review"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.status","published"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","2084"],["dc.bibliographiccitation.issue","8"],["dc.bibliographiccitation.journal","The EMBO Journal"],["dc.bibliographiccitation.lastpage","2091"],["dc.bibliographiccitation.volume","18"],["dc.contributor.author","Dierks, Thomas"],["dc.contributor.author","Lecca, M.Rita"],["dc.contributor.author","Schlotterhose, Petra"],["dc.contributor.author","Schmidt, Bernhard"],["dc.contributor.author","Figura, Kurt von"],["dc.date.accessioned","2019-07-10T08:12:46Z"],["dc.date.available","2019-07-10T08:12:46Z"],["dc.date.issued","1999"],["dc.description.abstract","Sulfatases carry at their catalytic site a unique posttranslational modification, an a-formylglycine residue that is essential for enzyme activity. Formylglycine is generated by oxidation of a conserved cysteine or, in some prokaryotic sulfatases, serine residue. In eukaryotes, this oxidation occurs in the endoplasmic reticulum during or shortly after import of the nascent sulfatase polypeptide. The modification of arylsulfatase A was studied in vitro and was found to be directed by a short linear sequence, CTPSR, starting with the cysteine to be modified. Mutational analyses showed that the cysteine, proline and arginine are the key residues within this motif, whereas formylglycine formation tolerated the individual, but not the simultaneous substitution of the threonine or serine. The CTPp. motif was transferred to a heterologous protein leading to low-efficient formylglycine formation. The efficiency reached control values when seven additional residues (AALLTGR) directly following the CTPSR motif in arylsulfatase A were present. Mutating up to four residues simultaneously within this heptamer sequence inhibited the modification only moderately. AALLTGR may, therefore, have an auxiliary function in presenting the core motif to the modifying enzyme. Within the two motifs, the key residues are fully, and other residues are highly conserved among all known members of the sulfatase family."],["dc.format.mimetype","application/pdf"],["dc.identifier.purl","https://resolver.sub.uni-goettingen.de/purl?goescholar/3445"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/61034"],["dc.language.iso","en"],["dc.notes.intern","Migrated from goescholar"],["dc.publisher","Nature Publishing Group"],["dc.relation.issn","0261-4189"],["dc.rights","Goescholar"],["dc.rights.access","openAccess"],["dc.rights.uri","https://goescholar.uni-goettingen.de/licenses"],["dc.subject","cysteine; endoplasmic reticulum; multiple sulfatase deficiency; protein modification; sulfatase"],["dc.subject.ddc","610"],["dc.title","Sequence determinants directing conversion of cysteine to formylglycine in eukaryotic sulfatases"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.version","published_version"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","5828"],["dc.bibliographiccitation.issue","8"],["dc.bibliographiccitation.journal","Journal of Biological Chemistry"],["dc.bibliographiccitation.lastpage","5839"],["dc.bibliographiccitation.volume","288"],["dc.contributor.author","Ennemann, Eva C."],["dc.contributor.author","Radhakrishnan, Karthikeyan"],["dc.contributor.author","Mariappan, Malaiyalam"],["dc.contributor.author","Wachs, Michaela"],["dc.contributor.author","Pringle, Thomas H."],["dc.contributor.author","Schmidt, Bernhard"],["dc.contributor.author","Dierks, Thomas"],["dc.date.accessioned","2018-11-07T09:28:01Z"],["dc.date.available","2018-11-07T09:28:01Z"],["dc.date.issued","2013"],["dc.description.abstract","Formylglycine-generating enzyme (FGE) post-translationally converts a specific cysteine in newly synthesized sulfatases to formylglycine (FGly). FGly is the key catalytic residue of the sulfatase family, comprising 17 nonredundant enzymes in human that play essential roles in development and homeostasis. FGE, a resident protein of the endoplasmic reticulum, is also secreted. A major fraction of secreted FGE is N-terminally truncated, lacking residues 34-72. Here we demonstrate that this truncated form is generated intracellularly by limited proteolysis mediated by proprotein convertase(s) (PCs) along the secretory pathway. The cleavage site is represented by the sequence RYSR72 down arrow, a motif that is conserved in higher eukaryotic FGEs, implying important functionality. Residues Arg-69 and Arg-72 are critical because their mutation abolishes FGE processing. Furthermore, residues Tyr-70 and Ser-71 confer an unusual property to the cleavage motif such that endogenous as well as overexpressed FGE is only partially processed. FGE is cleaved by furin, PACE4, and PC5a. Processing is disabled in furin-deficient cells but fully restored upon transient furin expression, indicating that furin is the major protease cleaving FGE. Processing by endogenous furin occurs mostly intracellularly, although also extracellular processing is observed in HEK293 cells. Interestingly, the truncated form of secreted FGE no longer possesses FGly-generating activity, whereas the unprocessed form of secreted FGE is active. As always both forms are secreted, we postulate that furin-mediated processing of FGE during secretion is a physiological means of higher eukaryotic cells to regulate FGE activity upon exit from the endoplasmic reticulum."],["dc.description.sponsorship","Deutsche Forschungsgemeinschaft [DI 575/7, SCHM 830/2, SFB 860]"],["dc.identifier.doi","10.1074/jbc.M112.405159"],["dc.identifier.isi","000315342500053"],["dc.identifier.pmid","23288839"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/30675"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Amer Soc Biochemistry Molecular Biology Inc"],["dc.relation.issn","0021-9258"],["dc.title","Proprotein Convertases Process and Thereby Inactivate Formylglycine-generating Enzyme"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.status","published"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","11556"],["dc.bibliographiccitation.issue","17"],["dc.bibliographiccitation.journal","Journal of Biological Chemistry"],["dc.bibliographiccitation.lastpage","11564"],["dc.bibliographiccitation.volume","283"],["dc.contributor.author","Mariappan, Malaiyalam"],["dc.contributor.author","Gande, Santosh Lakshmi"],["dc.contributor.author","Radhakrishnan, Karthikeyan"],["dc.contributor.author","Schmidt, Bernhard"],["dc.contributor.author","Dierks, Thomas"],["dc.contributor.author","von Figura, Kurt"],["dc.date.accessioned","2018-11-07T11:15:58Z"],["dc.date.available","2018-11-07T11:15:58Z"],["dc.date.issued","2008"],["dc.description.abstract","Formylglycine-generating enzyme (FGE) catalyzes the oxidation of a specific cysteine residue in nascent sulfatase polypeptides to formylglycine (FGly). This FGly is part of the active site of all sulfatases and is required for their catalytic activity. Here we demonstrate that residues 34-68 constitute an N-terminal extension of the FGE catalytic core that is dispensable for in vitro enzymatic activity of FGE but is required for its in vivo activity in the endoplasmic reticulum (ER), i.e. for generation of FGly residues in nascent sulfatases. In addition, this extension is needed for the retention of FGE in the ER. Fusing a KDEL retention signal to the C terminus of FGE is sufficient to mediate retention of an N-terminally truncated FGE but not sufficient to restore its biological activity. Fusion of FGE residues 1-88 to secretory proteins resulted in ER retention of the fusion protein. Moreover, when fused to the paralog of FGE (pFGE), which itself lacks FGly-generating activity, the FGE extension ( residues 34-88) of this hybrid construct led to partial restoration of the biological activity of co-expressed N-terminally truncated FGE. Within the FGE N-terminal extension cysteine 52 is critical for the biological activity. We postulate that this N-terminal region of FGE mediates the interaction with an ER component to be identified and that this interaction is required for both the generation of FGly residues in nascent sulfatase polypeptides and for retention of FGE in the ER."],["dc.identifier.doi","10.1074/jbc.M707858200"],["dc.identifier.isi","000255067400056"],["dc.identifier.pmid","18305113"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/54486"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Amer Soc Biochemistry Molecular Biology Inc"],["dc.relation.issn","0021-9258"],["dc.title","The non-catalytic N-terminal extension of formylglycine-generating enzyme is required for its biological activity and retention in the endoplasmic reticulum"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.status","published"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","1020"],["dc.bibliographiccitation.issue","9"],["dc.bibliographiccitation.journal","European Journal of Human Genetics"],["dc.bibliographiccitation.lastpage","1023"],["dc.bibliographiccitation.volume","21"],["dc.contributor.author","Schlotawa, Lars"],["dc.contributor.author","Radhakrishnan, Karthikeyan"],["dc.contributor.author","Baumgartner, Matthias"],["dc.contributor.author","Schmid, Regula"],["dc.contributor.author","Schmidt, Bernhard"],["dc.contributor.author","Dierks, Thomas"],["dc.contributor.author","Gärtner, Jutta"],["dc.date.accessioned","2017-09-07T11:47:37Z"],["dc.date.available","2017-09-07T11:47:37Z"],["dc.date.issued","2013"],["dc.description.abstract","Multiple sulfatase deficiency (MSD) is a rare inborn error of metabolism affecting posttranslational activation of sulfatases by the formylglycine generating enzyme (FGE). Due to mutations in the encoding SUMF1 gene, FGE's catalytic capacity is impaired resulting in reduced cellular sulfatase activities. Both, FGE protein stability and residual activity determine disease severity and have previously been correlated with the clinical MSD phenotype. Here, we report a patient with a late infantile severe course of disease. The patient is compound heterozygous for two so far undescribed SUMF1 mutations, c.156delC (p.C52fsX57) and c.390A>T (p.E130D). In patient fibroblasts, mRNA of the frameshift allele is undetectable. In contrast, the allele encoding FGE-E130D is expressed. FGE-E130D correctly localizes to the endoplasmic reticulum and has a very high residual molecular activity in vitro (55% of wildtype FGE); however, it is rapidly degraded. Thus, despite substantial residual enzyme activity, protein instability determines disease severity, which highlights that potential MSD treatment approaches should target protein folding and stabilization mechanisms."],["dc.identifier.doi","10.1038/ejhg.2012.291"],["dc.identifier.gro","3142298"],["dc.identifier.isi","000323281400021"],["dc.identifier.pmid","23321616"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/6731"],["dc.notes.intern","WoS Import 2017-03-10 / Funder: Deutsche Forschungsgemeinschaft; Fonds der Chemischen Industrie"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.publisher","Nature Publishing Group"],["dc.relation.issn","1018-4813"],["dc.title","Rapid degradation of an active formylglycine generating enzyme variant leads to a late infantile severe form of multiple sulfatase deficiency"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.journal","Journal of Inherited Metabolic Disease"],["dc.bibliographiccitation.volume","33"],["dc.contributor.author","Schlotawa, Lars"],["dc.contributor.author","Radhakrishnan, Karthikeyan"],["dc.contributor.author","Schmid, R."],["dc.contributor.author","Schmidt, B."],["dc.contributor.author","Dierks, Thomas"],["dc.contributor.author","Gaertner, J."],["dc.contributor.author","Baumgartner, M."],["dc.date.accessioned","2018-11-07T08:40:54Z"],["dc.date.available","2018-11-07T08:40:54Z"],["dc.date.issued","2010"],["dc.identifier.isi","000281735000463"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/19349"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Springer"],["dc.publisher.place","Dordrecht"],["dc.title","FUNCTIONAL CHARACTERIZATION OF TWO NOVEL SUMF1 MUTATIONS LEADING TO A MILD PHENOTYPE IN MULTIPLE SULFATASE DEFICIENCY"],["dc.type","conference_abstract"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","269"],["dc.bibliographiccitation.issue","2"],["dc.bibliographiccitation.journal","Journal of Molecular Biology"],["dc.bibliographiccitation.lastpage","277"],["dc.bibliographiccitation.volume","305"],["dc.contributor.author","Bülow, Rixa von"],["dc.contributor.author","Schmidt, Bernhard"],["dc.contributor.author","Dierks, Thomas"],["dc.contributor.author","Figura, Kurt von"],["dc.contributor.author","Usón, Isabel"],["dc.date.accessioned","2018-11-07T09:27:13Z"],["dc.date.available","2018-11-07T09:27:13Z"],["dc.date.issued","2001"],["dc.description.abstract","Arylsulfatase A (ASA) belongs to the sulfatase family whose members carry a C-alpha-formylglycine that is post-translationally generated by oxidation of a conserved cysteine or serine residue. The crystal structures of two arylsulfatases, ASA and ASB, and kinetic studies on ASA mutants led to different proposals for the catalytic mechanism in the hydrolysis of sulfate esters. The structures of two ASA mutants that lack the functional C-alpha-formylglycine residue 69, in complex with a synthetic substrate, have been determined in order to unravel the reaction mechanism. The crystal structure of the inactive mutant C69A-ASA in complex with p-nitrocatechol sulfate (pNCS) mimics a reaction intermediate during sulfate ester hydrolysis by the active enzyme, without the covalent bond to the key side-chain FGly69. The structure shows that the side-chains of lysine 123, lysine 302, serine 150, histidine 229, the main-chain of the key residue 69 and the divalent cation in the active center are involved in sulfate binding. It is proposed that histidine 229 protonates the leaving alcoholate after hydrolysis. C69S-ASA is able to bind covalently to the substrate and hydrolyze it, but is unable to release the resulting sulfate. Nevertheless, the resulting sulfation is low. The structure of C69S-ASA shows the serine side-chain in a single conformation, turned away from the position a substrate occupies in the complex. This suggests that the double conformation observed in the structure of wild-tips ASA is more likely to correspond to a formylglycine hydrate than to a twofold disordered aldehyde oxo group, and accounts for the relative inertness of the C69S-ASA mutant. In the C69S-ASA-pNCS complex, the substrate occupies the same position as in the C69A-ASA-pNCS complex, which corresponds to the noncovalently bonded substrate. Based on the structural data, a detailed mechanism for sulfate ester cleavage is proposed, involving an aldehyde hydrate as the functional group. (C) 2001 Academic Press."],["dc.identifier.doi","10.1006/jmbi.2000.4297"],["dc.identifier.isi","000166413600008"],["dc.identifier.pmid","11124905"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/30484"],["dc.language.iso","en"],["dc.notes.status","final"],["dc.notes.submitter","Najko"],["dc.relation.issn","0022-2836"],["dc.title","Crystal structure of an enzyme-substrate complex provides insight into the interaction between human arylsulfatase A and its substrates during catalysis"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","3262"],["dc.bibliographiccitation.issue","17"],["dc.bibliographiccitation.journal","FEBS Journal"],["dc.bibliographiccitation.lastpage","3274"],["dc.bibliographiccitation.volume","282"],["dc.contributor.author","Peng, Jianhe"],["dc.contributor.author","Alam, Sarfaraz"],["dc.contributor.author","Radhakrishnan, Karthikeyan"],["dc.contributor.author","Mariappan, Malaiyalam"],["dc.contributor.author","Rudolph, Markus Georg"],["dc.contributor.author","May, Caroline"],["dc.contributor.author","Dierks, Thomas"],["dc.contributor.author","von Figura, Kurt"],["dc.contributor.author","Schmidt, Bernhard"],["dc.date.accessioned","2018-11-07T09:52:38Z"],["dc.date.available","2018-11-07T09:52:38Z"],["dc.date.issued","2015"],["dc.description.abstract","C alpha-formylglycine (FGly) is the catalytic residue of sulfatases in eukaryotes. It is generated by a unique post-translational modification catalysed by the FGly-generating enzyme (FGE) in the endoplasmic reticulum. FGE oxidizes a cysteine residue within the conserved CxPxR sequence motif of nascent sulfatase polypeptides to FGly. Here we show that this oxidation is strictly dependent on molecular oxygen (O-2) and consumes 1 mol O-2 per mol FGly formed. For maximal activity FGE requires an O-2 concentration of 9% (105 mu M). Sustained FGE activity further requires the presence of a thiol-based reductant such as DTT. FGly is also formed in the absence of DTT, but its formation ceases rapidly. Thus inactivated FGE accumulates in which the cysteine pair Cys336/Cys341 in the catalytic site is oxidized to form disulfide bridges between either Cys336 and Cys341 or Cys341 and the CxPxR cysteine of the sulfatase. These results strongly suggest that the Cys336/Cys341 pair is directly involved in the O-2-dependent conversion of the CxPxR cysteine to FGly. The available data characterize eukaryotic FGE as a monooxygenase, in which Cys336/Cys341 disulfide bridge formation donates the electrons required to reduce one oxygen atom of O-2 to water while the other oxygen atom oxidizes the CxPxR cysteine to FGly. Regeneration of a reduced Cys336/Cys341 pair is accomplished in vivo by a yet unknown reductant of the endoplasmic reticulum or in vitro by DTT. Remarkably, this monooxygenase reaction utilizes O-2 without involvement of any activating cofactor."],["dc.identifier.doi","10.1111/febs.13347"],["dc.identifier.isi","000360629200002"],["dc.identifier.pmid","26077311"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/36170"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Wiley-blackwell"],["dc.relation.issn","1742-4658"],["dc.relation.issn","1742-464X"],["dc.title","Eukaryotic formylglycine-generating enzyme catalyses a monooxygenase type of reaction"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.status","published"],["dc.type.subtype","letter_note"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","9455"],["dc.bibliographiccitation.issue","11"],["dc.bibliographiccitation.journal","Journal of Biological Chemistry"],["dc.bibliographiccitation.lastpage","9461"],["dc.bibliographiccitation.volume","277"],["dc.contributor.author","Bülow, Rixa von"],["dc.contributor.author","Schmidt, Bernhard"],["dc.contributor.author","Dierks, Thomas"],["dc.contributor.author","Schwabauer, Nelli"],["dc.contributor.author","Schilling, Klaus"],["dc.contributor.author","Weber, Ekkehard"],["dc.contributor.author","Usón, Isabel"],["dc.contributor.author","Figura, Kurt von"],["dc.date.accessioned","2018-11-07T10:31:08Z"],["dc.date.available","2018-11-07T10:31:08Z"],["dc.date.issued","2002"],["dc.description.abstract","In one of the most common mutations causing metachromatic leukodystrophy, the P426L-allele of arylsulfatase A (ASA), the deficiency of ASA results from its instability in lysosomes. Inhibition of lysosomal cysteine proteinases protects the P426L-ASA and restores the sulfatide catabolism in fibroblasts of the patients. P426L-ASA, but not wild type ASA, was cleaved by purified cathepsin L at threonine 421 yielding 54- and 9-kDa fragments. X-ray crystallography at 2.5-Angstrom resolution showed that cleavage is not due to a difference in the protein fold that would expose the peptide bond following threonine 421 to proteases. Octamerization, which depends on protonation of Glu-424, was impaired for P426L-ASA. The mutation lowers the pH for the octamer/ dimer equilibrium by 0.6 pH units from pH 5.8 to 5.2. A second oligomerization mutant (ASA-A464R) was generated that failed to octamerize even at pH 4.8. A464R-ASA was degraded in lysosomes to catalytically active 54-kDa intermediate. In cathepsin L-deficient fibroblasts, degradation of P426L-ASA and A464R-ASA to the 54-kDa fragment was reduced, while further degradation was blocked. This indicates that defective oligomerization of ASA allows degradation of ASA to a catalytically active 54-kDa intermediate by lysosomal cysteine proteinases, including cathepsin L. Further degradation of the 54-kDa intermediate critically depends on cathepsin L and is modified by the structure of the 9-kDa cleavage product."],["dc.identifier.doi","10.1074/jbc.M111993200"],["dc.identifier.isi","000174400600096"],["dc.identifier.pmid","11777924"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/44031"],["dc.language.iso","en"],["dc.notes.status","final"],["dc.notes.submitter","Najko"],["dc.relation.issn","0021-9258"],["dc.title","Defective oligomerization of arylsulfatase A as a cause of its instability in lysosomes and metachromatic leukodystrophy"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","483"],["dc.bibliographiccitation.issue","6"],["dc.bibliographiccitation.journal","Structure"],["dc.bibliographiccitation.lastpage","491"],["dc.bibliographiccitation.volume","9"],["dc.contributor.author","Boltes, Imke"],["dc.contributor.author","Czapinska, Honorata"],["dc.contributor.author","Kahnert, Antje"],["dc.contributor.author","Bülow, Rixa von"],["dc.contributor.author","Dierks, Thomas"],["dc.contributor.author","Schmidt, Bernhard"],["dc.contributor.author","Figura, Kurt von"],["dc.contributor.author","Kertesz, Michael A."],["dc.contributor.author","Uson, Isabel"],["dc.date.accessioned","2018-11-07T09:02:54Z"],["dc.date.available","2018-11-07T09:02:54Z"],["dc.date.issued","2001"],["dc.description.abstract","Background: Sulfatases constitute a family of enzymes with a highly conserved active site region including a C alpha -formylglycine that is posttranslationally generated by the oxidation of a conserved cysteine or serine residue. The crystal structures of two human arylsulfatases, ASA and ASB, along with ASA mutants and their complexes led to different proposals for the catalytic mechanism in the hydrolysis of sulfate esters. Results: The crystal structure of a bacterial sulfatase from Pseudomonas aeruginosa (PAS) has been determined at 1.3 Angstrom. Fold and active site region are strikingly similar to those of the known human sulfatases. The structure allows a precise determination of the active site region, unequivocally showing the presence of a C alpha -formylglycine hydrate as the key catalytic residue. Furthermore, the cation located in the active site is unambiguously characterized as calcium by both its B value and the geometry of its coordination sphere. The active site contains a noncovalently bonded sulfate that occupies the same position as the one in para-nitrocate-cholsulfate in previously studied ASA complexes. Conclusions: The structure of PAS shows that the resting state of the key catalytic residue in sulfatases is a formylglycine hydrate. These structural data establish a mechanism for sulfate ester cleavage involving an aldehyde hydrate as the functional group that initiates the reaction through a nucleophilic attack on the sulfur atom in the substrate. The alcohol is eliminated from a reaction intermediate containing pentacoordinated sulfur. Subsequent elimination of the sulfate regenerates the aldehyde, which is again hydrated. The metal cation involved in stabilizing the charge and anchoring the substrate during catalysis is established as calcium."],["dc.identifier.doi","10.1016/S0969-2126(01)00609-8"],["dc.identifier.isi","000169240800004"],["dc.identifier.pmid","11435113"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/24780"],["dc.language.iso","en"],["dc.notes.status","final"],["dc.notes.submitter","Najko"],["dc.relation.issn","0969-2126"],["dc.title","1.3 Å Structure of Arylsulfatase from Pseudomonas aeruginosa Establishes the Catalytic Mechanism of Sulfate Ester Cleavage in the Sulfatase Family"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dspace.entity.type","Publication"]]