Identification and cloning of an aspartyl proteinase from Coccidioides immitis

Johnson, Suzanne M.; Kerekes, K. M.; Zimmermann, C. R.; Williams, Robert H.; Pappagianis, Demosthenes

doi:10.1016/s0378-1119(99)00478-3

Cited by 15 publications

(15 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The WoLF PSORT algorithm predicted Pep1 to be cell wall associated, secreted into the culture medium, or both. (23). The hydrophobicity profile of Pep1 predicted three hydrophobic domains (aa 180 to 222, 275 to 305, and 345 to 380).…”

Section: Vol 74 2006mentioning

confidence: 96%

“…Sequence matches were also manually verified. On this basis of peptide sequence matches we identified an open reading frame (ORF) of a 1.4-kb gene in the C. posadasii genome database which revealed 99.8% nucleotide sequence identity to the cDNA of a previously reported gene that encodes an aspartyl protease of Coccidioides (23). The genomic and cDNA sequences of the gene identified in the C. posadasii database, which were confirmed by cloning and nucleotide sequence analysis as described below, are designated in this paper as PEP1.…”

Section: Examinations Of the Incidence Of Coccidioidal Infections Inmentioning

confidence: 97%

“…A BLAST search of the Swiss-Prot/TrEMBL and nonredundant NCBI protein databases using the translated ORF identified an aspartyl protease of C. posadasii which had previously been described (GenBank accession no. AF162132) (23). The earlier reported gene (PEP) was isolated from the Silveira strain of C. posadasii.…”

Section: Vol 74 2006mentioning

confidence: 99%

See 2 more Smart Citations

A Recombinant Aspartyl Protease ofCoccidioides posadasiiInduces Protection against Pulmonary Coccidioidomycosis in Mice

et al. 2006

View full text Add to dashboard Cite

Coccidioidomycosis is a respiratory disease of humans caused by the desert soil-borne fungal pathogens Coccidioides spp. Recurrent epidemics of this mycosis in the southwestern United States have contributed significantly to escalated health care costs. Clinical and experimental studies indicate that prior symptomatic coccidioidomycosis induces immunity against subsequent infection, and activation of T cells is essential for containment of the pathogen and its clearance from host tissue. Development of a human vaccine against coccidioidomycosis has focused on recombinant T-cell-reactive antigens which elicit a durable protective immune response against pulmonary infection in mice. In this study we fractionated a protective multicomponent parasitic cell wall extract in an attempt to identify T-cell antigens. Immunoblots of electrophoretic separations of this extract identified patient seroreactive proteins which were subsequently excised from two-dimensional polyacrylamide gel electrophoresis gels, trypsin digested, and sequenced by tandem mass spectrometry. The full-length gene which encodes a dominant protein in the immunoblot was identified using established methods of bioinformatics. The gene was cloned and expressed, and the recombinant protein was shown to stimulate immune T cells in vitro. The deduced protein was predicted to contain epitopes that bind to human major histocompatibility complex class II molecules using a TEPITOPE-based algorithm. Synthetic peptides corresponding to the predicted T-cell epitopes induced gamma interferon production by immune T lymphocytes. The T-cell-reactive antigen, which is homologous to secreted fungal aspartyl proteases, protected mice against pulmonary infection with Coccidioides posadasii. We argue that this immunoproteomic/ bioinformatic approach to the identification of candidate vaccines against coccidioidomycosis is both efficient and productive.

show abstract

Section: Vol 74 2006mentioning

confidence: 96%

Section: Examinations Of the Incidence Of Coccidioidal Infections Inmentioning

confidence: 97%

See 1 more Smart Citation

A Recombinant Aspartyl Protease ofCoccidioides posadasiiInduces Protection against Pulmonary Coccidioidomycosis in Mice

et al. 2006

View full text Add to dashboard Cite

show abstract

“…An aspartic protease (Asp 1) of 45 kDa was identified from a soluble vaccine formulation, named F27K [122], prepared from formalin-killed spherules of C. immitis [123]. Similarly, the major proteinaceous component (Pep1 of 43 kDa) of a Triton X-114 detergent extraction from spherule wall fraction of the C. immitis possessed aspartic protease activity [124].…”

Section: Aspartic Proteasesmentioning

confidence: 99%

Aspartic Proteases of Human Pathogenic Fungi are Prospective Targets for the Generation of Novel and Effective Antifungal Inhibitors

Santos

2011

CEI

View full text Add to dashboard Cite

Fungi can cause life-threatening diseases, particularly in patients with weakened immunological systems. Although treatment options are available for these individuals, dose-limiting toxicity and the appearance of drug-resistant microorganisms are growing problems. Detailed structural and functional characterization of fungal proteases has led to novel insights into the workings of these fascinating catalytic machines. Identification and characterization of proteasemediated processes in human pathogenic fungi is progressing at a rapid rate. In these microorganisms, aspartic-type proteases carry out "housekeeping" tasks common to many eukaryotes as well as functions highly specific to the fungal life cycles. Consequently, the possibility of developing selective inhibitors of key aspartic proteases of pathogenic fungi into novel chemotherapeutic strategies is being vigorously explored. The present review describes the knowledge in the area of aspartic proteases produced by human fungal pathogens. As well, the effects of aspartic proteolytic inhibitors on multiple vital processes of fungal cells, with special emphasis on their roles to arrest fungal development and virulence, will be presented and discussed.

show abstract

“…1): the first seeks to provide fractions and then to dissect out the protective components; the second seeks to identify, e.g., by amino acid sequence, specific proteins for the cloning of DNA and the preparation of recombinant proteins. One such component, aspartyl protease, has been cloned by Johnson in our laboratory (Johnson et al, 1999).…”

Section: Immune Protection-vaccine Studiesmentioning

confidence: 99%

Identification and cloning of an aspartyl proteinase from Coccidioides immitis

Cited by 15 publications

References 26 publications

A Recombinant Aspartyl Protease ofCoccidioides posadasiiInduces Protection against Pulmonary Coccidioidomycosis in Mice

A Recombinant Aspartyl Protease ofCoccidioides posadasiiInduces Protection against Pulmonary Coccidioidomycosis in Mice

Aspartic Proteases of Human Pathogenic Fungi are Prospective Targets for the Generation of Novel and Effective Antifungal Inhibitors

Seeking a Vaccine against Coccidioides immitis and Serologic Studies: Expectations and Realities

Contact Info

Product

Resources

About