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Updated: Apr 16, 2026
Separation and Fractionation of Cell Wall and Cell Membrane Proteins from Mycobacterium tuberculosis for Downstream Protein Analysis
Published on: September 26, 2025
[Proteomic analysis of U937 cells expressing Mycobacterium tuberculosis heat shock protein 16.3]
Taiming Li1, Xingyuan Yang, Qinglin Meng
1College of Life Science and Technology, China Pharmaceutical University, Nanjing 215163, China.
Objective:
To compare the protein expression differences between U937 macrophages expressing M. tuberculosis (MTB) Hsp16.3 protein and U937 macrophages expressing green fluorescent protein (GFP), and therefore to explore the protein expressions related to latent TB infection(LTBI).
Methods:
U937 macrophages were infected with an integrase-deficient Lentivirus vector to transiently express MTB Hsp16.3, and green fluorescent protein (GFP) as a control. 2-DE was used to compare the differentially expressed proteins in the infected U937 cells. Then 5 significantly different expressed protein spots were identified by using mass spectrometry.
Results:
The data of 6 protein spots in gel obtained from peptide mass fingerprinting were retrieved in protein database. They were identified as heat shock protein 70, actin, elongation factor I, peptidyl-prolyl cis-trans isomerase, ubiquitin-conjugation enzyme E2, and milk acyl glutathione lyase.
Conclusion:
The results showed that MTB specific protein intrusion resulted in changes of macrophage proteome expression, and this finding may help in understanding of the interaction between macrophages and MTB specific proteins.
