Chakrabarti Amrita, Kaushik Monika, Khan Juveria, Soota Deepanshu, Ponnusamy Kalairasan, Saini Sunil, Manvati Siddharth, Singhal Jhalak, Ranganathan Anand, Pati Soumya, Dhar Pawan Kumar, Singh Shailja
Department of Life Sciences, Shiv Nadar University, Greater Noida 201314, Uttar Pradesh, India.
Special Centre for Molecular Medicine, Jawaharlal Nehru University, New Delhi 110067, India.
ACS Omega. 2022 May 25;7(22):18361-18373. doi: 10.1021/acsomega.2c00661. eCollection 2022 Jun 7.
We asked if transfer RNA (tRNA) ever got an opportunity of translating its own sequence during evolution, what would have been the function of such tRNA-encoded peptides (tREPs)? If not, could one artificially synthesize tREPs to study the corresponding functional outcomes? Here, we report a novel, first-in-the-class, chemically synthesized tREP-18 molecule originating from the tRNA sequence showing potent antileishmanial property. As a first step, tRNAs were computationally translated into peptide sequence equivalents and a database of full-length hypothetical tREPs was created. The tREP sequences were sent into sequence, structure, and energy filters to narrow down potential peptides for experimental validation. Based on the functional predictions, tREPs were screened against antiparasitic targets, leading to the identification of tREP-18 as a potential antiparasitic peptide. The in vitro assay of chemically synthesized tREP-18 on the strain of showed its potent antileishmanial property (IC50 value of 22.13 nM). The atomic force microscopy and scanning electron microscopy images indicated significant alteration in the cytoskeletal architecture of tREP-18-treated parasites. Also, tREP-18 seems to destabilize the mitochondrial membrane potential of parasites, disrupting their cellular integrity and leading to parasitic death. The cellular assays of the tREP-18 peptide on the BS12 strain, a clinical isolate of post-kala azar dermal leishmaniasis, demonstrated its significant efficacy at an IC50 value of 15 nM. The tREP-18 peptide showed a toxic effect on the amastigote stage of the parasite, showing macrophage pathogen clearance at a concentration of 22.5 nM. This study provides the proof of the concept of making a new class of functional peptides from tRNA sequences. It also opens a huge untapped tRNA-peptide space toward novel discoveries and applications. In the future, it would be interesting to perform tREP edits and redesign tREPs toward specific applications.
在进化过程中，如果转运核糖核酸（tRNA）曾有机会翻译其自身序列，那么这种由tRNA编码的肽（tREP）会有什么功能？如果没有，能否人工合成tREP来研究相应的功能结果？在此，我们报告了一种新型的、同类首创的化学合成tREP - 18分子，它源自tRNA序列，具有强大的抗利什曼原虫特性。第一步，通过计算机将tRNA翻译成等效的肽序列，并创建了一个全长假设tREP的数据库。将tREP序列输入序列、结构和能量筛选器，以缩小潜在肽段范围用于实验验证。基于功能预测，针对抗寄生虫靶点筛选tREP，从而鉴定出tREP - 18为一种潜在的抗寄生虫肽。对化学合成的tREP - 18进行体外检测，结果表明其对[具体菌株]具有强大的抗利什曼原虫特性（IC50值为22.13 nM）。原子力显微镜和扫描电子显微镜图像显示，经tREP - 18处理的寄生虫的细胞骨架结构发生了显著改变。此外，tREP - 18似乎会破坏寄生虫的线粒体膜电位，扰乱其细胞完整性并导致寄生虫死亡。对tREP - 18肽在黑热病后皮肤利什曼病临床分离株BS12菌株上进行细胞检测，结果表明其在IC50值为15 nM时具有显著疗效。tREP - 18肽对寄生虫的无鞭毛体阶段显示出毒性作用，在浓度为22.5 nM时可清除巨噬细胞中的病原体。这项研究为从tRNA序列制造一类新的功能性肽提供了概念验证。它还为新发现和应用开辟了一个巨大的未开发的tRNA - 肽空间。未来，对tREP进行编辑并针对特定应用重新设计tREP将会很有趣。
