title: Identification and characterization of an aspartyl protease from Cryptococcus neoformans.
abstract: Cryptococcosis, caused by Cryptococcus neoformans, is an invasive infection often occurring in AIDS patients. Potent therapy against HIV, which includes protease inhibitors (PIs), has beneficial effects also on opportunistic infections by pathogens such as C. neoformans and C. albicans. PIs inhibit growth of C. albicans by affecting the activity of its aspartyl proteases. We identified, cloned and sequenced a cDNA from C. neoformans encoding for a putative aspartyl protease (CnAP1), and the corresponding genomic region. The gene cnap1 codifies for a protein of 505 aa, with a canonical aspartyl protease structure. We purified the recombinant protein and analyzed its activity in the presence of PIs (Indinavir, Lopinavir, Ritonavir), but did not evidence any inhibition of protease activity. The transcriptional level of cnap1 in C. neoformans is constant in different media. The absence of any inhibition activity by PIs suggests that other targets for PIs might exist in C. neoformans.
authors.affiliation: Department of Biomedical Sciences, Chair of Immunology, University of Modena and Reggio Emilia, via Campi 287, 41100 Modena, Italy.
authors.forename: Marcello
authors.initials: M
authors.lastname: Pinti
authors.forename: Carlotta Francesca
authors.initials: CF
authors.lastname: Orsi
authors.forename: Lara
authors.initials: L
authors.lastname: Gibellini
authors.forename: Roberto
authors.initials: R
authors.lastname: Esposito
authors.forename: Andrea
authors.initials: A
authors.lastname: Cossarizza
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Blasi
authors.forename: Samuele
authors.initials: S
authors.lastname: Peppoloni
authors.forename: Cristina
authors.initials: C
authors.lastname: Mussini
mesh_terms: D016282:Aspartic Acid Endopeptidases / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D019295:Computational Biology; D003455:Cryptococcus neoformans / Q000201:enzymology* / Q000235:genetics; D004271:DNA, Fungal; D005800:Genes, Fungal; D007438:Introns; D008958:Models, Molecular; D016366:Open Reading Frames; D017434:Protein Structure, Tertiary; D017422:Sequence Analysis, DNA
chemical_list: D004271:DNA, Fungal; D016282:Aspartic Acid Endopeptidases