PE/PPE Proteome and ESX-5 Substrate Spectrum in Mycobacterium marinum
Abstract
:1. Introduction
2. Results
2.1. Proteinase AspN Treatment Improved the Coverage of PE/PPE Proteins
2.2. Quantification of PE/PPE Protein Abundance
2.3. EspG5-PE/PPE Interactome
2.4. Mapping PE/PPE Protein Networks
2.5. Interaction of EspG5 with PPE31 and PPE32
3. Discussion
4. Materials and Methods
4.1. Bacterial Strain and Growth Condition
4.2. Preparation of Culture Filtrate
4.3. Cloning and Purification of the Recombinant Proteins
4.4. Pulldown Assays
4.5. Protease Digestion
4.6. Mass Spectrometry
4.7. LC-MS Data Analysis
4.8. Western Analysis
4.9. Structural Model Prediction
4.10. Phylogenetic Tree and PE–PPE Pairs Calculation
4.11. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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PE | PE_PGRS | PPE | Total | |
---|---|---|---|---|
AspN unique | 3 | 18 | 6 | 27 |
Trypsin unique | 1 | 14 | 8 | 23 |
AspN/Trypsin | 9 | 39 | 38 | 86 |
Total (% PE/PPE) | 13 (48.1%) | 71 (47.8%) | 52 (49.1%) | 136 (48.4%) |
PE_ORF1 | PE_GENE1 | PPE_ORF2 | PPE_GENE2 | Coefficient | Sij | Rij |
---|---|---|---|---|---|---|
MMAR_3763 | PE_PGRS27 | MMAR_1950 | PPE15_1 | 0.99849 | 0.770717 | 0.673922 |
MMAR_1208 | PE_PGRS | MMAR_3666 | PPE40 | 0.998099 | 0.894426 | 0.588773 |
MMAR_3467 | PE | MMAR_1514 | PPE51_1 | 0.997299 | 0.775825 | 0.65997 |
MMAR_3316 | PE_PGRS | MMAR_3666 | PPE40 | 0.996912 | 0.887535 | 0.604463 |
MMAR_2010 | PE_PGRS | MMAR_2685 | PPE30 | 0.996684 | 0.797027 | 0.478634 |
MMAR_3467 | PE | MMAR_3444 | PPE61 | 0.996225 | 0.787156 | 0.495639 |
MMAR_2010 | PE_PGRS | MMAR_3666 | PPE40 | 0.988286 | 0.795013 | 0.582921 |
MMAR_2894 | PE | MMAR_5448 | PPE68 | 0.982382 | 0.974785 | 0.784697 |
MMAR_1208 | PE_PGRS | MMAR_2685 | PPE30 | 0.980908 | 0.89644 | 0.480968 |
MMAR_0370 | PE | MMAR_4218 | PPE | 0.976292 | 0.81602 | 0.605133 |
MMAR_3316 | PE_PGRS | MMAR_4319 | PPE13 | 0.966084 | 0.891999 | 0.35109 |
MMAR_0370 | PE | MMAR_3444 | PPE61 | 0.964489 | 0.906339 | 0.499527 |
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Yan, L.; Lai, H.Y.; Leung, T.C.N.; Cheng, H.F.; Chen, X.; Tsui, S.K.W.; Ngai, S.M.; Au, S.W.N. PE/PPE Proteome and ESX-5 Substrate Spectrum in Mycobacterium marinum. Int. J. Mol. Sci. 2024, 25, 9550. https://doi.org/10.3390/ijms25179550
Yan L, Lai HY, Leung TCN, Cheng HF, Chen X, Tsui SKW, Ngai SM, Au SWN. PE/PPE Proteome and ESX-5 Substrate Spectrum in Mycobacterium marinum. International Journal of Molecular Sciences. 2024; 25(17):9550. https://doi.org/10.3390/ijms25179550
Chicago/Turabian StyleYan, Lili, Hiu Ying Lai, Thomas Chun Ning Leung, Hiu Fu Cheng, Xin Chen, Stephen Kwok Wing Tsui, Sai Ming Ngai, and Shannon Wing Ngor Au. 2024. "PE/PPE Proteome and ESX-5 Substrate Spectrum in Mycobacterium marinum" International Journal of Molecular Sciences 25, no. 17: 9550. https://doi.org/10.3390/ijms25179550