Reichard, Utz

[["dc.bibliographiccitation.firstpage","405"],["dc.bibliographiccitation.issue","1"],["dc.bibliographiccitation.journal","Infection and Immunity"],["dc.bibliographiccitation.lastpage","412"],["dc.bibliographiccitation.volume","69"],["dc.contributor.author","Zaugg, C."],["dc.contributor.author","Borg-von Zepelin, Margarete"],["dc.contributor.author","Reichard, Utz"],["dc.contributor.author","Sanglard, D."],["dc.contributor.author","Monod, Michel"],["dc.date.accessioned","2018-11-07T09:34:03Z"],["dc.date.available","2018-11-07T09:34:03Z"],["dc.date.issued","2001"],["dc.description.abstract","Medically important yeasts of the genus Candida secrete aspartic proteinases (Saps), which are of particular interest as virulence factors. Like Candida albicans, Candida tropicalis secretes in vitro one dominant Sap (Sapt1p) in a medium containing bovine serum albumin (BSA) as the sole source of nitrogen. Using the gene SAPT1 as a probe and under low-stringency hybridization conditions, three new closely related gene sequences, SAPT2 to SAPT4, encoding secreted proteinases were cloned from a C. tropicalis lambda EMBL3 genomic library. All bands identified by Southern blotting of EcoRI-digested C. tropicalis genomic DNA with SAPT1 could be assigned to a specific SAP gene. Therefore, the SAPT gene family of C. tropicalis is likely to contain only four members. Interestingly, the SAPT2 and SAPT3 gene products, Sapt2p and Sapt3p, which have not yet been detected in C. tropicalis cultures in vitro, were produced as active recombinant enzymes with the methylotrophic yeast Pichia pastoris as an expression system. As expected, reverse transcriptase PCR experiments revealed a strong SAPT1 signal with RNA extracted from cells grown in BSA medium. However, a weak signal was obtained with all other SAPT genes under several renditions tested, showing that these SAPT genes could be expressed at a basic level. Together, these experiments suggest that the gene products Sapt2p, Sapt3p, and Sapt4p could be produced under conditions yet to be described in vitro or during infection."],["dc.identifier.doi","10.1128/IAI.69.1.405-412.2001"],["dc.identifier.isi","000165943500050"],["dc.identifier.pmid","11119531"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/32096"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Amer Soc Microbiology"],["dc.relation.issn","0019-9567"],["dc.title","Secreted aspartic proteinase family of Candida tropicalis"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.status","published"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","954"],["dc.bibliographiccitation.issue","4"],["dc.bibliographiccitation.journal","Journal of Proteome Research"],["dc.bibliographiccitation.lastpage","962"],["dc.bibliographiccitation.volume","5"],["dc.contributor.author","Asif, Abdul Rahman"],["dc.contributor.author","Oellerich, M."],["dc.contributor.author","Amstrong, V. W."],["dc.contributor.author","Riemenschneider, B."],["dc.contributor.author","Monod, Michel"],["dc.contributor.author","Reichard, Utz"],["dc.date.accessioned","2018-11-07T10:02:59Z"],["dc.date.available","2018-11-07T10:02:59Z"],["dc.date.issued","2006"],["dc.description.abstract","Aspergillus fumigatus is a mold causing most of the invasive fungal lung infections in the immunocompromised host. In addition, the species is the causative agent of certain allergic diseases. Both in invasive and in allergic diseases, the conidial surface mediates the first contact with the human immune system. Thus, conidial surface proteins may be reasonable vaccine candidates as well as important allergens. To broaden the list of those antigens, intact viable Aspergillus conidia were extracted with mild alkaline buffer at pH 8.5 in the presence of a 1,3-beta-glucanase. The proteome of this fraction was separated by two- dimensional gel electrophoresis (2-DE) and analyzed by liquid chromatography coupled with tandem mass spectrometry. Altogether 26 different A. fumigatus proteins were identified, twelve of which contain a signal for secretion. Among these were the known major conidial surface protein rodlet A, one acid protease PEP2, one lipase, a putative disulfide isomerase and a putative fructose-1,6-biphosphatase. The known allergen Aspf 3 was identified among the proteins without a signal for secretion. On the basis of the recently annotated A. fumigatus genome (Nature 2005, 438, 1151-1156), proteome analysis is now a powerful tool to confirm expression of hypothetical proteins and, thereby to identify additional vaccine candidates and possible new allergens of this important fungal pathogen."],["dc.description.sponsorship","NIAID NIH HHS [U01 AI 48830]; Wellcome Trust"],["dc.identifier.doi","10.1021/pr0504586"],["dc.identifier.isi","000236816100025"],["dc.identifier.pmid","16602703"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/38345"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Amer Chemical Soc"],["dc.relation.issn","1535-3893"],["dc.title","Proteome of conidial surface associated proteins of Aspergillus fumigatus reflecting potential vaccine candidates and allergens"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.status","published"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","79"],["dc.bibliographiccitation.journal","Gene"],["dc.bibliographiccitation.lastpage","88"],["dc.bibliographiccitation.volume","339"],["dc.contributor.author","Jousson, Olivier"],["dc.contributor.author","Lechenne, B."],["dc.contributor.author","Bontems, O."],["dc.contributor.author","Mignon, B."],["dc.contributor.author","Reichard, Utz"],["dc.contributor.author","Barblan, J."],["dc.contributor.author","Quadroni, Manfredo"],["dc.contributor.author","Monod, Michel"],["dc.date.accessioned","2018-11-07T10:45:34Z"],["dc.date.available","2018-11-07T10:45:34Z"],["dc.date.issued","2004"],["dc.description.abstract","Secreted proteases constitute potential virulence factors of dermatophytes. A total of seven genes encoding putative serine proteases of the subtilisin family (SUB) were isolated in Trichophyton rubrum. Based on sequence data and intron-exon structure, a phylogenetic analysis of subtilisins from T rubrum and other fungi revealed a presumed ancestral lineage comprising T rubrum SUB2 and Aspergillus SUBs. All other SUBs (SUB1, SUB3-7) are dermatophyte-specific and have apparently emerged more recently, through successive gene duplication events. We showed that two subtilisins, Sub3 and Sub4, were detected in culture supernatants of T rubrum grown in a medium containing soy protein as a sole nitrogen source. Both recombinant enzymes produced in Pichia pastoris are highly active on keratin azure suggesting that these proteases play an important role in invasion of keratinised tissues by the fungus. The set of deduced amino acid sequences of T rubrum SUB ORFs allowed the identification of orthologous Subs secreted by other dermatophyte species using proteolysis and mass spectrometry. (C) 2004 Elsevier B.V. All rights reserved."],["dc.identifier.doi","10.1016/j.gene.2004.06.024"],["dc.identifier.isi","000224173200009"],["dc.identifier.pmid","15363848"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/47530"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.relation.issn","0378-1119"],["dc.title","Secreted subtilisin gene family in Trichophyton rubrum"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","87"],["dc.bibliographiccitation.lastpage","106"],["dc.contributor.author","Monod, Michel"],["dc.contributor.author","Jousson, Olivier"],["dc.contributor.author","Reichard, Utz"],["dc.contributor.editor","Latgé, Jean-Paul"],["dc.contributor.editor","Steinbach, William J."],["dc.date.accessioned","2021-12-08T12:28:05Z"],["dc.date.available","2021-12-08T12:28:05Z"],["dc.date.issued","2008"],["dc.identifier.doi","10.1128/9781555815523.ch8"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/95549"],["dc.language.iso","en"],["dc.notes.intern","DOI-Import GROB-476"],["dc.publisher","ASM Press"],["dc.publisher.place","Washington, DC, USA"],["dc.relation.eisbn","9781683671381"],["dc.relation.eisbn","9781555815523"],["dc.relation.isbn","9781555814380"],["dc.relation.ispartof","Aspergillus fumigatus\n and Aspergillosis"],["dc.title","Aspergillus fumigatus Secreted Proteases"],["dc.type","book_chapter"],["dc.type.internalPublication","yes"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","1541"],["dc.bibliographiccitation.journal","Microbiology"],["dc.bibliographiccitation.lastpage","1550"],["dc.bibliographiccitation.volume","157"],["dc.contributor.author","Sriranganadane, Dev"],["dc.contributor.author","Reichard, Utz"],["dc.contributor.author","Salamin, Karine"],["dc.contributor.author","Fratti, Marina"],["dc.contributor.author","Jousson, Olivier"],["dc.contributor.author","Waridel, Patrice"],["dc.contributor.author","Quadroni, Manfredo"],["dc.contributor.author","Neuhaus, Jean-Marc"],["dc.contributor.author","Monod, Michel"],["dc.date.accessioned","2018-11-07T08:56:19Z"],["dc.date.available","2018-11-07T08:56:19Z"],["dc.date.issued","2011"],["dc.description.abstract","In an acidic protein medium Aspergillus fumigatus secretes an aspartic endoprotease (Pep) as well as tripeptidyl-peptidases, a prolyl-peptidase and carboxypeptidases. In addition, LC-MS/MS revealed a novel glutamic protease, AfuGprA, homologous to Aspergillus niger aspergillopepsin II. The importance of AfuGprA in protein digestion was evaluated by deletion of its encoding gene in A. fumigatus wild-type D141 and in a pepA mutant. Either A. fumigatus Pep or AfuGprA was shown to be necessary for fungal growth in protein medium at low pH. Exoproteolytic activity is therefore not sufficient for complete protein hydrolysis and fungal growth in a medium containing proteins as the sole nitrogen source. Pep and AfuGprA constitute a pair of endoproteases active at low pH, in analogy to A. fumigatus alkaline protease (Alp) and metalloprotease I (Mep), where at least one of these enzymes is necessary for fungal growth in protein medium at neutral pH. Heterologous expression of AfuGprA in Pichia pastoris showed that the enzyme is synthesized as a preproprotein and that the propeptide is removed through an autoproteolytic reaction at low pH to generate the mature protease. In contrast to A. niger aspergillopepsin II, AfuGprA is a single-chain protein and is structurally more similar to G1 proteases characterized in other non-Aspergillus fungi."],["dc.description.sponsorship","Swiss National Foundation for Scientific Research [320030-1179641]"],["dc.identifier.doi","10.1099/mic.0.048603-0"],["dc.identifier.isi","000291179900029"],["dc.identifier.pmid","21349972"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/23119"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Soc General Microbiology"],["dc.relation.issn","1350-0872"],["dc.title","Secreted glutamic protease rescues aspartic protease Pep deficiency in Aspergillus fumigatus during growth in acidic protein medium"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.status","published"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","549"],["dc.bibliographiccitation.issue","6"],["dc.bibliographiccitation.journal","International Journal of Medical Microbiology"],["dc.bibliographiccitation.lastpage","558"],["dc.bibliographiccitation.volume","290"],["dc.contributor.author","Reichard, Utz"],["dc.contributor.author","Cole, G. T."],["dc.contributor.author","Hill, T. W."],["dc.contributor.author","Ruchel, R."],["dc.contributor.author","Monod, Michel"],["dc.date.accessioned","2018-11-07T09:39:36Z"],["dc.date.available","2018-11-07T09:39:36Z"],["dc.date.issued","2000"],["dc.description.abstract","A novel subtilisin-related serine proteinase (ALP2) [EC 3.4.21.48] with a broad range of activity between pH 4.5 and 11.0 was released from a cell wall fraction of Aspergillus fumigatus by an alkaline pH shift. The enzyme which was not detected in the culture supernatant was partially purified by phenylbutylamine agarose chromatography. The N-terminal sequence revealed that ALP2 is the same protein identified as the major allergen of A. fumigatus in patients suffering from extrinsic bronchial asthma (Shen et al. 1999, Int. Arch. Allergy Immunol. 119, 259-264). Based on this N-terminal sequence and on a conserved region of fungal subtilisins, a specific PCR probe was generated and the ALP2 genomic and cDNA were isolated from corresponding phage libraries. ALP2 shares a 49 % identity with the vacuolar proteinase B (PrB) of Saccharomyces cerevisiae. In addition there is a 78 % identity with PEPC, a serine proteinase which has been described in Aspergillus niger. Targeted disruption of the ALP2-encoding gene resulted in a slightly decreased speed of vegetative growth and in a more than 80% reduction of sporulation in the alp2-negative mutants, correlated with an approximately 50 % reduction of the median diameter of conidiophore vesicles. The requirement of ALP2 for regular sporulation, in addition to its role in allergic asthma, raises further interest in cellular proteinases in respect to morphogenesis and pathogenesis in A. fumigatus."],["dc.identifier.isi","000165333400007"],["dc.identifier.pmid","11100830"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/33322"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Urban & Fischer Verlag"],["dc.relation.issn","1438-4221"],["dc.title","Molecular characterization and influence on fungal development of ALP2, a novel serine proteinase from Aspergillus fumigatus"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.status","published"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.issue","5"],["dc.bibliographiccitation.journal","Mycoses"],["dc.bibliographiccitation.volume","53"],["dc.contributor.author","Lababidi, B."],["dc.contributor.author","Bader, Oliver"],["dc.contributor.author","Reichard, Utz"],["dc.contributor.author","Monod, Michel"],["dc.contributor.author","Sulahian, A."],["dc.contributor.author","Binder, L."],["dc.contributor.author","Gross, U."],["dc.contributor.author","Weig, M. S."],["dc.date.accessioned","2018-11-07T08:40:00Z"],["dc.date.available","2018-11-07T08:40:00Z"],["dc.date.issued","2010"],["dc.format.extent","400"],["dc.identifier.isi","000280998400044"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/19130"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Wiley-blackwell"],["dc.publisher.place","Malden"],["dc.relation.issn","0933-7407"],["dc.title","Evaluation of the specific IgG- and IgA-antibody response against immundominant Aspergillus fumigatus antigens as diagnostic markers of invasive aspergillosis"],["dc.type","conference_abstract"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.status","published"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","4704"],["dc.bibliographiccitation.issue","8"],["dc.bibliographiccitation.journal","Infection and Immunity"],["dc.bibliographiccitation.lastpage","4713"],["dc.bibliographiccitation.volume","73"],["dc.contributor.author","Denikus, N."],["dc.contributor.author","Orfaniotou, F."],["dc.contributor.author","Wulf, Gerald"],["dc.contributor.author","Lehmann, P. F."],["dc.contributor.author","Monod, Michel"],["dc.contributor.author","Reichard, Utz"],["dc.date.accessioned","2018-11-07T09:37:21Z"],["dc.date.available","2018-11-07T09:37:21Z"],["dc.date.issued","2005"],["dc.description.abstract","Rabbits that had been infected intravenously with conidiospores of Aspergillus fumigatus were used as sources of antibody for screening a X phage cDNA expression library. The cDNA was derived from A. fumigatus mRNA that had been extracted from newly formed, germling hyphae. Thirty-six antigens were identified using antisera from six rabbits. Though many of these antigens were expected to be intracellular proteins because their genes did not encode a signal sequence, the antisera showed consistently a stronger immunoblot reaction with a cell fraction enriched for the fungal cell wall than with a fraction of predominantly intracellular components. Antibodies to eight antigens, including the glycosylhydrolase Asp f 16, were produced by more than one rabbit. In current vaccine studies, Asp 116 is the only single antigen which has been reported to be capable of inducing protection against invasive aspergillosis in mice (S. Bozza et al., Microb. Infect. 4:1281-1290, 2002). Enolase and Aspergillus HSP90 were detected also; their homologues in Candida albicans have been tested as vaccines and have been reported to provide a partially protective response against invasive candidiasis in mice. The Aspergillus antigens reported here may have value both in diagnostic tests for different forms of aspergillosis and as vaccine candidates for protection against invasive disease."],["dc.description.sponsorship","NIAID NIH HHS [U01 AI 48830]; Wellcome Trust"],["dc.identifier.doi","10.1128/IAI.73.8.4704-4713.2005"],["dc.identifier.isi","000230760600028"],["dc.identifier.pmid","16040983"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/32823"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.relation.issn","0019-9567"],["dc.title","Fungal antigens expressed during invasive Aspergillosis"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","85"],["dc.bibliographiccitation.issue","1"],["dc.bibliographiccitation.journal","International Journal of Medical Microbiology"],["dc.bibliographiccitation.lastpage","96"],["dc.bibliographiccitation.volume","290"],["dc.contributor.author","Reichard, Utz"],["dc.contributor.author","Cole, G. T."],["dc.contributor.author","Ruchel, R."],["dc.contributor.author","Monod, Michel"],["dc.date.accessioned","2018-11-07T09:15:08Z"],["dc.date.available","2018-11-07T09:15:08Z"],["dc.date.issued","2000"],["dc.description.abstract","An aspartic proteinase PEP2 [EC 3.4.23.25] was purified from a cell wall fraction of Aspergillus fumigatus. The enzyme, which showed a broad range of activity from pH 2.0 to 7.0 and migrated as a single band of 39 kDa in SDS-PAGE, was not detected in the culture supernatant. A specific gene probe was designed on the basis of the N-terminal sequence of the native protein, and the PEP2 genomic and cDNA were isolated from corresponding libraries. The deduced amino acid sequence of PEP2 consists of 398 amino acids. A signal sequence of 18 amino acids and a proregion of another 52 amino acids were identified. The mature protein consists of 328 amino acids which include the two DTG-motifs of the active site common to almost all pepsin-like enzymes; PEP2 showed a 64% identity with the vacuolar proteinase A (PrA), of Saccharomyces cerevisiae, and an 88% identity with PEPE, an aspartic proteinase of Aspergillus niger. Recombinant PEP2 was overexpressed in Pichia pastoris and the active enzyme was secreted into the culture supernatant. Targeted deletion of PEP2 did not affect vegetative growth or cell and colony morphology. Identification of proteinases, such as PEP2, which are apparently associated with the Aspergillus cell wall raises new interest in these molecules with respect to their possible function in the pathogenesis of invasive aspergillosis."],["dc.identifier.isi","000086862900010"],["dc.identifier.pmid","11043985"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/27603"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Urban & Fischer Verlag"],["dc.relation.issn","1438-4221"],["dc.title","Molecular cloning and targeted deletion of PEP2 which encodes a novel aspartic proteinase from Aspergillus fumigatus"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.status","published"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","61"],["dc.bibliographiccitation.issue","1"],["dc.bibliographiccitation.journal","Medical Mycology"],["dc.bibliographiccitation.lastpage","71"],["dc.bibliographiccitation.volume","40"],["dc.contributor.author","Schoen, C."],["dc.contributor.author","Reichard, Utz"],["dc.contributor.author","Monod, Michel"],["dc.contributor.author","Kratzin, H. D."],["dc.contributor.author","Ruchel, R."],["dc.date.accessioned","2018-11-07T10:32:18Z"],["dc.date.available","2018-11-07T10:32:18Z"],["dc.date.issued","2002"],["dc.description.abstract","An extracellular aspartic proteinase (Rmap) from Rhizopus microsporus var. rhizopodiformis was detected in the culture supernatant of a fungal isolate from a case of rhinocerebral mucormycosis (case HA). The proteinase was purified to near homogeneity by ion exchange and affinity chromatography on pepstatin agarose. Based on its N-terminus the RMAP gene was cloned and found to code for 388 amino acids. The preproenzyme has an aminoterminal leader sequence of 65 amino acids, whereas the mature enzyme consists of 323 amino acids. The deduced amino-acid sequence of the preproenzyme was 82% homologous to an extracellular aspartic proteinase of Rhizopus niveus. Low stringency Southern blot analysis of R. microsporus DNA suggested the presence of other homologous genes. Expression of Rmap in Pichia pastoris was achieved, and the recombinant enzyme was active in the yeast culture supernatant. Both enzyme preparations exhibited a similar optimum of activity in the pH 2.5 region. Furthermore, Rmap was shown to activate bovine blood coagulation factor X at slightly acidic pH in vitro. Expression of the proteinase during mycosis was proven by a specific immune response of patient HA."],["dc.identifier.doi","10.1080/mmy.40.1.61.71"],["dc.identifier.isi","000173949300009"],["dc.identifier.pmid","11860014"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/44317"],["dc.language.iso","en"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.relation.issn","1369-3786"],["dc.title","Molecular cloning of an extracellular aspartic proteinase from Rhizopus microsporus and evidence for its expression during infection"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dspace.entity.type","Publication"]]