Comparative Proteomic Analysis of Cell Wall Proteins of Aminoglycosides Resistant and Sensitive Mycobacterium tuberculosis Clinical Isolates.

IF 1.9 4区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Current protein & peptide science Pub Date : 2024-11-07 DOI:10.2174/0113892037334796240927055243

Devesh Sharma, Sakshi Gautam, Nalini Srivastava, Abdul Mabood Khan, Deepa Bisht

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引用次数: 0

Abstract

Introduction: The rising prevalence of Mycobacterium tuberculosis (M.tb) strains resistant to aminoglycosides (amikacin and kanamycin) challenges effective TB control and treatment. Understanding the mechanisms behind this resistance is crucial since aminoglycosides are a mainstay of TB therapy.

Aim: The study aimed to analyze the cell wall proteins overexpressed in aminoglycoside-resistant isolates of Mycobacterium tuberculosis using proteomics approaches.

Methods: We used two-dimensional electrophoresis and mass spectrometry to compare the cell wall proteomes of aminoglycosides-resistant and susceptible clinical isolates. The overexpressed protein spots were excised and identified using liquid chromatography-mass spectrometry (LC/MS). The identified proteins were subsequently analyzed for molecular docking, pupylation site identification, and STRING analysis.

Results: We found a total of nine significantly upregulated proteins in aminoglycosides-resistant isolates. Three of these proteins were the same (isoform), resulting in the identification of seven unique proteins. Specifically, Rv3841 and Rv1308 belonged to intermediary metabolism and respiration; Rv2115c to the cell wall and cell processes; Rv2501c, Rv2247 and Rv0295c to lipid metabolism; and Rv2416c to virulence, detoxification/adaptation. Notably, variations in these proteins support cell wall integrity, aiding mycobacteria's establishment and proliferation. Molecular docking study revealed that both drugs bind strongly to the proteins' active site regions. Additionally, the GPS-PUP algorithm successfully identified possible pupylation sites within these proteins, except Rv0295c. Based on interactome analysis using the STRING 12.0 database, we have identified potential interactive partners suggesting their role in aminoglycosides resistance.

Conclusion: Overexpressed proteins not only act to counteract or regulate drug effects but also have a role in protein dynamics that allow for resistance. Some of these identified proteins may serve as innovative drug targets and biomarkers for the early detection of drug-specific resistance in M.tb. Further research is needed to elucidate the mechanisms by which these potential protein targets contribute to resistance in AK and KM M.tb isolates.

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对氨基糖苷类药物耐药和敏感的结核分枝杆菌临床分离株细胞壁蛋白质的比较蛋白质组学分析

导言：对氨基糖苷类药物（阿米卡星和卡那霉素）耐药的结核分枝杆菌（M.tb）菌株的流行率不断上升，给有效控制和治疗结核病带来了挑战。目的：本研究旨在利用蛋白质组学方法分析耐氨基糖苷类药物的结核分枝杆菌分离株中过度表达的细胞壁蛋白：我们使用二维电泳和质谱技术比较了耐氨糖苷类药物和易感的临床分离株的细胞壁蛋白质组。利用液相色谱-质谱法（LC/MS）切除过表达蛋白点并进行鉴定。随后对鉴定出的蛋白质进行了分子对接、蛹化位点鉴定和 STRING 分析：结果：我们在氨基糖苷类耐药分离物中发现了九种明显上调的蛋白质。结果：我们在氨基糖苷类耐药分离物中共发现了 9 个明显上调的蛋白质，其中 3 个蛋白质是相同的（异构体），因此鉴定出了 7 个独特的蛋白质。具体来说，Rv3841 和 Rv1308 属于中间代谢和呼吸；Rv2115c 属于细胞壁和细胞过程；Rv2501c、Rv2247 和 Rv0295c 属于脂质代谢；Rv2416c 属于毒力、解毒/适应。值得注意的是，这些蛋白质的变异支持细胞壁的完整性，有助于分枝杆菌的建立和增殖。分子对接研究表明，这两种药物都能与蛋白质的活性位点区域紧密结合。此外，除 Rv0295c 外，GPS-PUP 算法还成功识别了这些蛋白质中可能的蛹化位点。根据使用 STRING 12.0 数据库进行的相互作用组分析，我们确定了潜在的相互作用伙伴，表明它们在氨基糖苷类耐药性中的作用：结论：过度表达的蛋白质不仅能抵消或调节药物作用，还能在蛋白质动力学中发挥作用，从而产生抗药性。其中一些已发现的蛋白质可作为创新药物靶点和生物标志物，用于早期检测 M.tb 的特异性耐药性。要阐明这些潜在蛋白靶点在 AK 和 KM M.tb 分离物中产生抗药性的机制，还需要进一步的研究。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Current protein & peptide science 生物-生化与分子生物学

CiteScore

5.20

自引率

0.00%

发文量

审稿时长

6 months

期刊介绍： Current Protein & Peptide Science publishes full-length/mini review articles on specific aspects involving proteins, peptides, and interactions between the enzymes, the binding interactions of hormones and their receptors; the properties of transcription factors and other molecules that regulate gene expression; the reactions leading to the immune response; the process of signal transduction; the structure and function of proteins involved in the cytoskeleton and molecular motors; the properties of membrane channels and transporters; and the generation and storage of metabolic energy. In addition, reviews of experimental studies of protein folding and design are given special emphasis. Manuscripts submitted to Current Protein and Peptide Science should cover a field by discussing research from the leading laboratories in a field and should pose questions for future studies. Original papers, research articles and letter articles/short communications are not considered for publication in Current Protein & Peptide Science.