Molecular cloning of an extracellular aspartic proteinase from Rhizopus microsporus and evidence for its expression during infection

Détails

ID Serval
serval:BIB_3EB3E8F6C738
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Molecular cloning of an extracellular aspartic proteinase from Rhizopus microsporus and evidence for its expression during infection
Périodique
Medical Mycology
Auteur(s)
Schoen  C., Reichard  U., Monod  M., Kratzin  H. D., Ruchel  R.
ISSN
1369-3786 (Print)
Statut éditorial
Publié
Date de publication
02/2002
Volume
40
Numéro
1
Pages
61-71
Notes
Journal Article --- Old month value: Feb
Résumé
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.
Mots-clé
Amino Acid Sequence Animals Aspartic Endopeptidases/chemistry/*genetics/isolation & purification Cloning, Molecular Guinea Pigs Humans Molecular Sequence Data Mucormycosis/*enzymology Rhizopus/*enzymology
Pubmed
Web of science
Création de la notice
25/01/2008 16:46
Dernière modification de la notice
20/08/2019 13:35
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