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Proteomics

Proteomics

A proteome is the entire set of proteins that a cell type produces. We can study proteomes using the knowledge of genomes because genes code for mRNAs, and the mRNAs encode proteins. Although mRNA analysis is a step in the right direction, not all mRNAs are translated into proteins.
Proteomics is the study of proteomes' function. It involves the large-scale systematic study of the proteome to denote the protein complement expressed by a genome. Scientist Mark Wilkins coined the term...

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Related Experiment Video

Updated: Dec 1, 2025

Label-Free Quantitative Proteomics Workflow for Discovery-Driven Host-Pathogen Interactions

Label-Free Quantitative Proteomics Workflow for Discovery-Driven Host-Pathogen Interactions

Published on: October 20, 2020

Label-Free Quantitative Proteomics Workflow for Discovery-Driven Host-Pathogen Interactions.

Brianna Ball¹, Arjun Sukumaran¹, Jennifer Geddes-McAlister²

¹Molecular and Cellular Biology Department, University of Guelph.

Journal of Visualized Experiments : Jove

|November 9, 2020

Summary

This study introduces a label-free quantification (LFQ) method using mass spectrometry to analyze the proteome of the fungal pathogen Cryptococcus neoformans during host cell infection. The technique reveals key proteins involved in virulence and host response.

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Last Updated: Dec 1, 2025

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Published on: January 10, 2025

Area of Science:

Proteomics
Infectious Disease Research
Microbiology

Background:

Mass spectrometry (MS)-based quantitative proteomics enables global proteome analysis under various conditions.
Studying host-pathogen interactions is crucial for understanding infection dynamics.
Cryptococcus neoformans is a fungal pathogen causing cryptococcosis.

Purpose of the Study:

To develop and detail a label-free quantification (LFQ) workflow for analyzing the infectome of Cryptococcus neoformans interacting with host macrophage cells.
To provide a comprehensive characterization of both pathogen and host protein perspectives during infection.
To optimize protein preparation for simultaneous analysis of microbial and mammalian cells.

Main Methods:

Label-free quantification (LFQ) proteomics using liquid-chromatography coupled with tandem mass spectrometry (LC-MS/MS).
Standardized protein extraction protocols for both fungal pathogen (C. neoformans) and mammalian host cells (macrophages).
High-throughput, sensitive analysis of protein abundance profiles.

Main Results:

The LFQ method successfully quantified a wide dynamic range of proteins from both C. neoformans and host cells in a single experiment.
The workflow is transferable to various host-pathogen infection models.
Identified specific C. neoformans proteins crucial for virulence and host proteins responding to fungal invasion.

Conclusions:

The described LFQ workflow provides an unbiased and sensitive method for studying host-pathogen interactions.
This approach yields essential insights into microbial pathogenesis and host immune responses.
The protocol is adaptable for diverse infection settings, enhancing infectious disease research.