Identification of Serine 119 as an Effective Inhibitor Binding Site of M. tuberculosis Ubiquitin-like Protein Ligase PafA Using Purified Proteins and M. smegmatis
| العنوان: | Identification of Serine 119 as an Effective Inhibitor Binding Site of M. tuberculosis Ubiquitin-like Protein Ligase PafA Using Purified Proteins and M. smegmatis |
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| المؤلفون: | Fan-Lin Wu, Lijun Bi, Tao Wang, Jiao-Yu Deng, Jin Xie, Wang Xude, Shu-Juan Guo, Yang Li, Xian-En Zhang, He-wei Jiang, Zhaowei Xu, Daniel M. Czajkowsky, Jian-Feng Pei, Cheng-Xi Liu, Xiang He, Jiabin Wang, Sheng-Ce Tao, Hai-nan Zhang, Hong Chen |
| المصدر: | EBioMedicine, Vol 30, Iss, Pp 225-236 (2018) EBioMedicine |
| بيانات النشر: | Elsevier, 2018. |
| سنة النشر: | 2018 |
| مصطلحات موضوعية: | 0301 basic medicine, PafA, Cell Survival, Nitrogen, Ubiquitin-Protein Ligases, Mycobacterium smegmatis, lcsh:Medicine, 010402 general chemistry, medicine.disease_cause, 01 natural sciences, General Biochemistry, Genetics and Molecular Biology, Cell Line, Serine, Structure-Activity Relationship, 03 medical and health sciences, chemistry.chemical_compound, Bacterial Proteins, Ubiquitin, AEBSF, medicine, Humans, Amino Acid Sequence, Sulfones, Enzyme Inhibitors, Binding site, chemistry.chemical_classification, Mutation, DNA ligase, lcsh:R5-920, Binding Sites, M. Tuberculosis, Pupylation, biology, Chemistry, Macrophages, lcsh:R, 4-(2-aminoethyl) benzenesulfonyl fluoride, Mycobacterium tuberculosis, General Medicine, 0104 chemical sciences, Amino acid, Multiple drug resistance, 030104 developmental biology, Biochemistry, biology.protein, lcsh:Medicine (General), Research Paper |
| الوصف: | Owing to the spread of multidrug resistance (MDR) and extensive drug resistance (XDR), there is a pressing need to identify potential targets for the development of more-effective anti-M. tuberculosis (Mtb) drugs. PafA, as the sole Prokaryotic Ubiquitin-like Protein ligase in the Pup-proteasome System (PPS) of Mtb, is an attractive drug target. Here, we show that the activity of purified Mtb PafA is significantly inhibited upon the association of AEBSF (4-(2-aminoethyl) benzenesulfonyl fluoride) to PafA residue Serine 119 (S119). Mutation of S119 to amino acids that resemble AEBSF has similar inhibitory effects on the activity of purified Mtb PafA. Structural analysis reveals that although S119 is distant from the PafA catalytic site, it is located at a critical position in the groove where PafA binds the C-terminal region of Pup. Phenotypic studies demonstrate that S119 plays critical roles in the function of Mtb PafA when tested in M. smegmatis. Our study suggests that targeting S119 is a promising direction for developing an inhibitor of M. tuberculosis PafA. Highlights • The pupylation activity of purified M. tuberculosis PafA is almost completely inhibited upon the association of AEBSF. • The AEBSF binding site, Ser 119 plays critical roles in both the pupylation and depupylation activity of purified M. tuberculosis PafA. • Disruption of purified M. tuberculosis PafA Ser 119 causes a dramatic reduction in Pup binding. Drug-resistant tuberculosis is a major challenge worldwide, there is an urgent need to identify potential drug targets for developing more effective anti-tubercular drugs. M. tuberculosis ubiquitin-like protein ligase PafA is an attractive drug target, however, effective PafA inhibitors have not yet been identified. Here, we show that interruption of a single amino acid, S119, causes dramatic loss of PafA activity. S119 could thus serve as a promising precise target for developing M. tuberculosis PafA inhibitors. |
| اللغة: | English |
| تدمد: | 2352-3964 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::cc2f0477082b1f131ee07a99aad66bedTest http://www.sciencedirect.com/science/article/pii/S2352396418301129Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....cc2f0477082b1f131ee07a99aad66bed |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 23523964 |
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