Project ID: IPD3310

Project Title: Secretome analysis of Candida glabrata aspartyl protease-deficient strains

Principal Investigator: Dr Rupinder Kaur

PI Affiliation: Laboratory of Fungal Pathogenesis, Centre for DNA Fingerprinting and Diagnostics (CDFD), Hyderabad, India

Submitting Author: Dr Rupinder Kaur

Description: The project is aimed at characterizing the effect of deletion of surface associated aspartyl protease(s) on the secretome of of the human opportunistic fungal pathogen Candida glabrata.

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Meta Data

Sample Preparation: The supernatants of YNB medium-grown logarithmic-phase cultures of Cgyps7Δ and Cgyps2ΔypsCΔ strains of C. glabrata cells were collected in duplicates and passed through 0.4 µm membrane or syringe filters to remove the residual cells. The resultant filtrates were concentrated using Amicon Ultra-15 and Ultra-0.5 (10 kDa cutoff) centrifugal filter units. The concentrated secretome fractions were run on a 12% SDS-PAGE and stained with Coomassie Brilliant Blue. The gel lane containing all proteins was sliced into three sections, with each section containing a different size range (<50 kDa, 50-120 kDa and >120 kDa) of proteins.

Peptide Separation: Proteins were identified, after in-gel trypsin digestion, via the microcapillary LC-MS/MS (Liquid chromatography-tandem mass spectrometry) method at the Taplin Biological Mass Spectrometry Facility, Harvard Medical School, Boston, USA using the the LTQ Orbitrap Velos Pro ion-trap mass spectrometer.

Protein Characterization: The acquired fragmentation pattern for each peptide was analysed using the Sequest software, and searches were run against the UniProt C. glabrata reference proteome database. The identified peptides were filtered to 1% false discovery rate.

Experiment Type: Shotgun proteomics

PubMed-ID: 31621333

Species: Nakaseomyces glabratus-5478

Tissue:

Cell Type:

Disease: Unknown

Instrument Details: LTQ Orbitrap Velos (MS:1001742)

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