Hit Compounds and Associated Targets in Intracellular Mycobacterium tuberculosis
Abstract
:1. Introduction
2. Results and Discussion
2.1. Identification of Hit Compounds
2.2. Generation of Mutants, Mutant Characterization, and WGS
2.3. Identification of Candidate Intracellular Drug Targets in Resistant Mutants
3. Materials and Methods
3.1. Preparation of Chemical Compounds
3.2. Libraries
3.3. HTS
3.4. Bacterial Culture and Mutant Conditions
3.5. WGS of Mutants
3.6. Bioinformatics Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
Sample Availability
References
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ID | Molecular Weight | Intracellular MIC90 (μM) | Resistant Mutant Selection Media | Number of Resistant Colonies |
---|---|---|---|---|
213A | 365 | 0.6 | 5× MIC glucose | 5 |
267A | 433 | 0.16 | 2× MIC90 ADC | 3 |
290A | 476 | 0.6 | 2× MIC90 ADS | 2 |
950A | 436 | 1.26 | 2× MIC90 ADC | 4 |
739A | 381 | 2 | 2× MIC90 acetate | 3 |
472A | 355 | 2 | 2× MIC90 acetate | 4 |
412A | 380 | 0.32 | 2× MIC90 ADC | 3 |
412A | 380 | 0.32 | 2× MIC90 glucose | 6 |
296A | 420 | 1 | 2× MIC90 acetate | 6 |
648X | 386 | 1.58 | 2× MIC90 ADC | 2 |
454A | 496 | 2 | 2× MIC90 ADC | 3 |
1114A | 347 | 2.51 | 5× MIC90 glucose | 2 |
486X | 220 | 0.16 | 5× MIC90 ADC | 3 |
912A | 394 | 1.58 | 2× MIC90 acetate | 4 |
622A | 288 | 2 | 5× MIC90 glucose | 3 |
705A | 310.8 | 1.26 | 2× MIC acetate | 1 |
Compound | Number of Mutants | Gene/ ORFs | Genetic Polymorphisms (Frequency) | Relevant Codon Change (Frequency) | Product |
---|---|---|---|---|---|
213A | 5 | mmpL3 | 755A > G; 758G > A; 875T > C; 1985C > A; 2051T > C | Tyr252Cys; Gly253Glu; Ile292Thr; Ala662Glu; Val684Ala | Conserved membrane transport protein |
sugl | 16C > T | Gln6*, gained stop codon (2) | Involved in transport of sugar across the membrane. Translocation of the substrate. | ||
267A | 3 | mmpL3 | 763T > C; 765C > G; 1932 C > A | Phe255Leu (2); Phe644Leu | Conserved membrane transport protein |
290A | 2 | mmpL3 | 1909C > A | Leu637Ile (2) | Conserved membrane transport protein |
Rv0370c | 474G > T | Val158Val (2) | Unknown. possible Oxidoreductase | ||
950A | 4 | rpoB | 1720G > A | Asp574Asn (4) | Transcription of DNA into RNA |
Rv3629 | 641G > A | Gly214Glu (4) | Probable conserved integral membrane protein | ||
739A | 3 | ethA | 611T > C | Met204Thr (2) | Monooxygenase that activates the pro-drug ethionamide (ETH) |
rpsO | 157C > T | Arg53Trp | 30S ribosomal protein S15 | ||
Rv1024 | 154C > T | Pro52Ser | Possible conserved membrane protein | ||
Rv3220c | 746C > CA (2) | indels, frameshift variant | Probable two-component sensor kinase | ||
472A | 4 | ethA | 611T > C | Met204Thr (3) | Monooxygenase that activates the pro-drug ETH |
rpsO | 157C > T | Arg53Trp | 30S ribosomal protein S15 | ||
Rv3220c | 746C > CA (3) | indels, frameshift variant | Probable two-component sensor kinase | ||
412A | 9 | prrB | 452T > C (7); 548C > T; 875A > G | Leu151Pro (7); Thr183Ile; Gln292Arg; | Two-component regulatory system PRRA/PRRB |
moaC3 | 392A > G | Asp131Gly | Probable molybdenum cofactor biosynthesis protein | ||
iniB | 290C > T | Thr97Ile | Isoniazid inductible gene protein. | ||
narL | 298G > C | Ala100Pro | Possible nitrate/nitrite response transcriptional regulatory protein | ||
ctpl | 3113TGCGAG > T | Indels, frameshift variant | Probable cation-transporter ATPase I | ||
296A | 6 | TB18.5 | 145G > C; 236A > G; 243C > A; 277A > G | Val49Leu (2); Tyr79Cys; His81Gln; Thr93Ala (2) | Conserved protein |
Rv1948 | 122C > A | Ala41Glu | Hypothetical protein | ||
pks6 | 667G > A | Val223Ile (2) | Probable membrane-bound polyketide synthase | ||
648X | 2 | ctpC | 1511C > T | Ser504Phe | Probable metal cation-transporting P-type ATPase C |
454A | 3 | Rv0678 | 466 G > GC (3) | indels, frameshift variation | Conserved protein |
mbtA | 1369CT > C (3) | Indels, frameshift variation | Bifunctional enzyme MbtA: salicyl-AMP ligase (SAL-AMP ligase) + salicyl-S-ArCP synthetase | ||
1114A | 2 | dnaE1 | 2215A > G | Met739Val | Probable DNA polymerase III (alpha chain) DnaE1 (DNA nucleotidyltransferase) |
virS | 983C > A | Pro328His | Virulence-regulating transcriptional regulator VirS (AraC/XylS family) | ||
Rv0585c | 1202A > C | Asp401Ala | Conserved integral membrane protein | ||
sugl | 16C > T | Gln6* (2), gained stop codon | Involved in transport of sugar across the membrane. Responsible for the translocation of the substrate across the membrane. | ||
486X | 3 | phoR | 661G > C | Ala221Pro, | Possible two-component system response sensor kinase membrane associated PhoR |
fbiC | 1082C > A | Thr361Lys, | Probable F420 biosynthesis protein FbiC | ||
fbiA | 866T > A | Leu289Gln (2) | Probable F420 biosynthesis protein FbiA | ||
Rv3327 | 296C > G | Pro100Ala | Probable transposase fusion protein | ||
912A | 3 | ethA | 205T > C; 190T > C | Trp69Arg; Phe64Ile | Monooxygenase that activates the pro-drug ETH |
Rv2542 | 1042G > A | Ala348Thr (2) | Conserved hypothetical protein | ||
Rv3083 | 783G > A; 806T > C | Trp261*, gained stop codon; Leu269Pro | Probable monooxygenase (hydroxylase) | ||
622A | 3 | ftsK | 1192T > C | Ser398Pro | Possible cell division transmembrane protein |
virS | 712G > T | Val238Phe | Virulence-regulating transcriptional regulator VirS (AraC/XylS family) | ||
sugl | 16C > T | Gln6* (3), gained stop codon | Involved in transport of sugar across the membrane. Responsible for the translocation of the substrate across the membrane. | ||
705A | 1 | Rv3083 | 380_381 G > GA, indels, frame shift variant | Glu127_Thr128fs | Probable monooxygenase (hydroxylase) |
sugl | 16C > T | Gln6*, gained stop codon | Involved in transport of sugar across the membrane. Responsible for the translocation of the substrate across the membrane. |
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Tsui, C.K.M.; Sorrentino, F.; Narula, G.; Mendoza-Losana, A.; del Rio, R.G.; Herrán, E.P.; Lopez, A.; Bojang, A.; Zheng, X.; Remuiñán-Blanco, M.J.; et al. Hit Compounds and Associated Targets in Intracellular Mycobacterium tuberculosis. Molecules 2022, 27, 4446. https://doi.org/10.3390/molecules27144446
Tsui CKM, Sorrentino F, Narula G, Mendoza-Losana A, del Rio RG, Herrán EP, Lopez A, Bojang A, Zheng X, Remuiñán-Blanco MJ, et al. Hit Compounds and Associated Targets in Intracellular Mycobacterium tuberculosis. Molecules. 2022; 27(14):4446. https://doi.org/10.3390/molecules27144446
Chicago/Turabian StyleTsui, Clement K. M., Flavia Sorrentino, Gagandeep Narula, Alfonso Mendoza-Losana, Ruben Gonzalez del Rio, Esther Pérez Herrán, Abraham Lopez, Adama Bojang, Xingji Zheng, Modesto Jesus Remuiñán-Blanco, and et al. 2022. "Hit Compounds and Associated Targets in Intracellular Mycobacterium tuberculosis" Molecules 27, no. 14: 4446. https://doi.org/10.3390/molecules27144446