Variability of Aspergillus nidulans antigens with media and time and temperature of growth

Calera, José Antonio; López-Medrano, Ramiro; Ovejero, M. C.; Puente, Pilar Herrera; Leal, Fernando

doi:10.1128/iai.62.6.2322-2333.1994

Cited by 18 publications

(17 citation statements)

References 18 publications

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“…As shown in our previous studies (4), which describe the effect of the culture media used on the presence of Aspergillus antigens specifically reactive to aspergilloma sera, CDA medium induced a strong biosynthesis of immunodominant antigens. The proteins present in total mycelial extracts from A. nidulans grown at 37ЊC in CDA medium for 8 days were separated by SDS-PAGE (16% polyacrylamide) and silver stained or electrotransferred to nitrocellulose papers.…”

Section: Identification Of a Nidulans Antigens Consistently Recognizsupporting

confidence: 59%

“…Electrophoretic blotting procedures and immunological detection of proteins. Proteins from extracts were first subjected to electrophoresis (SDS-PAGE or two-dimensional gel electrophoresis) and then transferred to nitrocellulose sheets (0.45 m pore diameter; Schleicher & Schuell) in a Trans-Blot Cell (Bio-Rad) as previously described (4). The blots were soaked for 15 min in 10 mM Tris-HCl-150 mM NaCl (pH 7.5) (TNa) and then were soaked in 5% nonfat dry milk in 20 mM Tris-HCl (pH 7.5)-500 mM NaCl (TBS) for 1 h at room temperature to saturate additional protein-binding sites.…”

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confidence: 99%

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Characterization of the Aspergillus nidulans aspnd1 gene demonstrates that the ASPND1 antigen, which it encodes, and several Aspergillus fumigatus immunodominant antigens belong to the same family

et al. 1997

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For the first time, an immunodominant Aspergillus nidulans antigen (ASPND1) consistently reactive with serum samples from aspergilloma patients has been purified and characterized, and its coding gene (aspnd1) has been cloned and sequenced. ASPND1 is a glycoprotein with four N-glycosidically-bound sugar chains (around 2.1 kDa each) which are not necessary for reactivity with immune human sera. The polypeptide part is synthesized as a 277-amino-acid precursor of 30.6 kDa that after cleavage of a putative signal peptide of 16 amino acids, affords a mature protein of 261 amino acids with a molecular mass of 29 kDa and a pI of 4.24 (as deduced from the sequence). The ASPND1 protein is 53.1% identical to the AspfII allergen from Aspergillus fumigatus and 48% identical to an unpublished Candida albicans antigen. All of the cysteine residues and most of the glycosylation sites are perfectly conserved in the three proteins, suggesting a similar but yet unknown function. Analysis of the primary structure of the ASPND1 coding gene (aspnd1) has allowed the establishment of a clear relationship between several previously reported A. fumigatus and A. nidulans immunodominant antigens. The increasing incidence of Aspergillus-related diseases (43) has prompted many investigators to search for fungal molecules relevant either for the immunodiagnosis of or for understanding the pathology of the different forms of aspergillosis (ranging from pulmonary aspergilloma to invasive aspergillosis, passing through allergic situations). The role of several putative virulence factors has been investigated without too much success (for a review, see reference 3). In contrast, several antigens of demonstrated value as immunodiagnostic reagents for different Aspergillus infections have been purified, and some of their coding genes have been cloned. The AspfI ribotoxin (27), the AspfII allergen (2), and heat shock protein 1 (HSP-1) (16) from Aspergillus fumigatus have been obtained as recombinant antigens. The CAT1 (the subunit of a catalase) (22) and superoxide dismutase (13) antigens have been purified to homogeneity from A. fumigatus water-soluble extracts. All of these antigens have been shown to be consistently reactive with sera from patients with different forms of aspergillosis but not with sera from control or healthy individuals (2, 10, 24, 26). Certain other purified molecules, such as the 58-kDa (6) and gp55 (40) antigens, or semipurified fractions, such as the CS2 complex (30) or the cytosolic fraction complex (CFC) (24) from A. fumigatus, have been claimed to be useful reagents for immunodiagnosis. In this paper, we attempt to simplify the entangled antigenic array of A. fumigatus antigens by using data obtained from the purification and immunochemical characterization of a previously reported Aspergillus nidulans antigen (4) (now designated ASPND1), cloning of its coding gene, and analysis of its primary structure. The conclusion of this study is that several previously reported antigens or antigenic Aspergillus preparations conta...

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Section: Identification Of a Nidulans Antigens Consistently Recognizsupporting

confidence: 59%

mentioning

confidence: 99%

Characterization of the Aspergillus nidulans aspnd1 gene demonstrates that the ASPND1 antigen, which it encodes, and several Aspergillus fumigatus immunodominant antigens belong to the same family

et al. 1997

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“…A. nidulans TLK12 ( pabaA1, yA2 , Δ argB::trpC ::Δ B , Δ catB, trpC801, veA1 ) was obtained from J. Aguirre, Mexico City, Mexico [4]. Organisms were maintained, grown and harvested as described elsewhere [7,8]. Water‐soluble fractions (WSF) and cell culture supernatants (CCS) were obtained as previously described [7,8].…”

Section: Methodsmentioning

confidence: 99%

“…Organisms were maintained, grown and harvested as described elsewhere [7,8]. Water‐soluble fractions (WSF) and cell culture supernatants (CCS) were obtained as previously described [7,8].…”

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confidence: 99%

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Distinctive properties of the catalase B of Aspergillus nidulans

et al. 2000

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Aspergillus nidulans catalase B (CatB) was purified to homogeneity and characterized as a hydroperoxidase which resembles typical catalases in some physicochemical characteristics: (1) it has an apparent molecular weight of 360 000 and is composed of four glycosylated subunits, (2) it has hydrophobic properties as revealed by extractability in ethanol/chloroform and binding to phenyl-Superose, and (3) it has an acidic isoelectric point at pH 3.5. Also CatB exhibits some distinctive properties, e.g. it is not inhibited by the presence of 2% sodium dodecyl sulfate, 9 M urea or reducing agents. Furthermore, even though CatB does not exhibit any residual peroxidase activity, it is able to retain up to 38% of its initial catalase activity after incubation with the typical catalase inhibitor 3-amino-1,2,4-triazole. ß

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Biochemistry of Enzymatic Pathogenicity Factors

Cole

1996

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Variability of Aspergillus nidulans antigens with media and time and temperature of growth

Cited by 18 publications

References 18 publications

Characterization of the Aspergillus nidulans aspnd1 gene demonstrates that the ASPND1 antigen, which it encodes, and several Aspergillus fumigatus immunodominant antigens belong to the same family

Characterization of the Aspergillus nidulans aspnd1 gene demonstrates that the ASPND1 antigen, which it encodes, and several Aspergillus fumigatus immunodominant antigens belong to the same family

Distinctive properties of the catalase B of Aspergillus nidulans

Biochemistry of Enzymatic Pathogenicity Factors

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