Global profiling of secreted protease activities in
6 affiliations
- 1Center for Discovery and Innovation in Parasitic Diseases, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California San Diego, La Jolla, CA, USA
- 2Department of Molecular and Cell Biology, University of California Merced, Merced, CA, USA; Quantitative and Systems Biology Graduate Program, University of California Merced, Merced, CA, USA; Health Sciences Research Institute, University of California Merced, Merced, CA, USA
- 3Department of Molecular and Cell Biology, University of California Merced, Merced, CA, USA; Quantitative and Systems Biology Graduate Program, University of California Merced, Merced, CA, USA
- 4Department of Molecular and Cell Biology, University of California Merced, Merced, CA, USA
- 5Department of Molecular and Cell Biology, University of California Merced, Merced, CA, USA; Health Sciences Research Institute, University of California Merced, Merced, CA, USA; Center for Cellular and Biomolecular Machines, University of California Merced, Merced, CA, USA; Corresponding author at: Department of Molecular and Cell Biology, University of California Merced, Merced, CA, USA.
- 6Center for Discovery and Innovation in Parasitic Diseases, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California San Diego, La Jolla, CA, USA; Corresponding author at: Center for Discovery and Innovation in Parasitic Diseases, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California San Diego, La Jolla, CA, USA.
Abstract
Coccidioidomycosis (Valley fever), caused by inhalation of Coccidioides arthroconidia, remains a diagnostic challenge due to its clinical similarity to other respiratory illnesses and the absence of a rapid, definitive diagnostic test. Current approaches, including serology and imaging, often result in delayed or inconclusive diagnosis. Here, we profiled secreted protease activity in cell-free conditioned media from a single strain of Coccidioides posadasii . This activity could potentially form the basis of a protease-targeted diagnostic assay for clinical specimens. Using multiplex substrate profiling by mass spectrometry (MSP-MS) applied to cell-free conditioned media, we characterized global proteolytic activity at acidic (pH 2.5) and neutral (pH 7.4) conditions. At acidic pH, we detected dominant aspartyl protease activity that was inhibited by pepstatin and exhibited substrate preferences similar to human cathepsin D. Consistent with these findings, a commercial cathepsin D fluorogenic substrate was efficiently cleaved under acidic conditions. At neutral pH, we observed distinct proteolytic activity consistent with contributions from both exopeptidases and endopeptidases. Inhibition profiling and fluorogenic substrate assays further suggested involvement of serine and threonine proteases. These findings provide an activity-based characterization of secreted proteases from a single C. posadasii strain and establish a resource for future studies evaluating their biological and diagnostic relevance.
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BibTeX
@article{serrano2026global,
title = {{Global profiling of secreted protease activities in}},
author = {Ana K. Serrano and Tahirah Williams and Austin M. Perry and Mélanie A.C. Ikeh and Tvisha Devavarapu and Pavla Fajtova and Zhenze Jiang and Clarissa J. Nobile and Anthony J. O’Donoghue},
journal = {Current Research in Microbial Sciences},
year = {2026},
month = oct,
doi = {10.1016/j.crmicr.2026.100671},
url = {https://doi.org/10.1016/j.crmicr.2026.100671},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Global profiling of secreted protease activities in
AU - Ana K. Serrano
AU - Tahirah Williams
AU - Austin M. Perry
AU - Mélanie A.C. Ikeh
AU - Tvisha Devavarapu
AU - Pavla Fajtova
AU - Zhenze Jiang
AU - Clarissa J. Nobile
AU - Anthony J. O’Donoghue
PY - 2026
DA - 2026/10/17
JO - Current Research in Microbial Sciences
DO - 10.1016/j.crmicr.2026.100671
UR - https://doi.org/10.1016/j.crmicr.2026.100671
AB - Coccidioidomycosis (Valley fever), caused by inhalation of Coccidioides arthroconidia, remains a diagnostic challenge due to its clinical similarity to other respiratory illnesses and the absence of a rapid, definitive diagnostic test. Current approaches, including serology and imaging, often result in delayed or inconclusive diagnosis. Here, we profiled secreted protease activity in cell-free conditioned media from a single strain of Coccidioides posadasii . This activity could potentially form the basis of a protease-targeted diagnostic assay for clinical specimens. Using multiplex substrate profiling by mass spectrometry (MSP-MS) applied to cell-free conditioned media, we characterized global proteolytic activity at acidic (pH 2.5) and neutral (pH 7.4) conditions. At acidic pH, we detected dominant aspartyl protease activity that was inhibited by pepstatin and exhibited substrate preferences similar to human cathepsin D. Consistent with these findings, a commercial cathepsin D fluorogenic substrate was efficiently cleaved under acidic conditions. At neutral pH, we observed distinct proteolytic activity consistent with contributions from both exopeptidases and endopeptidases. Inhibition profiling and fluorogenic substrate assays further suggested involvement of serine and threonine proteases. These findings provide an activity-based characterization of secreted proteases from a single C. posadasii strain and establish a resource for future studies evaluating their biological and diagnostic relevance.
SN - 2666-5174
ER -