protein conformation analysis

  • 文章类型: Journal Article
    蛋白质结构测定在深度学习模型的帮助下取得了进展,能够从蛋白质序列中预测蛋白质折叠。然而,在蛋白质结构仍未描述的某些情况下,获得准确的预测变得至关重要。这在处理稀有时尤其具有挑战性,多样的结构和复杂的样品制备。不同的指标评估预测可靠性,并提供对结果强度的洞察,通过结合不同的模型,提供对蛋白质结构的全面了解。在之前的研究中,研究了两种名为ARM58和ARM56的蛋白质。这些蛋白质包含四个功能未知的结构域,存在于利什曼原虫中。ARM是指抗锑标记物。这项研究的主要目的是评估模型预测的准确性,从而提供对这些发现背后的复杂性和支持指标的见解。该分析还扩展到从其他物种和生物体获得的预测的比较。值得注意的是,这些蛋白质中的一种与克氏锥虫和布鲁氏锥虫具有直系同源物,对我们的分析有进一步的意义。这一尝试强调了评估深度学习模型的不同输出的重要性。促进不同生物体和蛋白质之间的比较。这在没有先前结构信息可用的情况下变得特别相关。
    Protein structure determination has made progress with the aid of deep learning models, enabling the prediction of protein folding from protein sequences. However, obtaining accurate predictions becomes essential in certain cases where the protein structure remains undescribed. This is particularly challenging when dealing with rare, diverse structures and complex sample preparation. Different metrics assess prediction reliability and offer insights into result strength, providing a comprehensive understanding of protein structure by combining different models. In a previous study, two proteins named ARM58 and ARM56 were investigated. These proteins contain four domains of unknown function and are present in Leishmania spp. ARM refers to an antimony resistance marker. The study\'s main objective is to assess the accuracy of the model\'s predictions, thereby providing insights into the complexities and supporting metrics underlying these findings. The analysis also extends to the comparison of predictions obtained from other species and organisms. Notably, one of these proteins shares an ortholog with Trypanosoma cruzi and Trypanosoma brucei, leading further significance to our analysis. This attempt underscored the importance of evaluating the diverse outputs from deep learning models, facilitating comparisons across different organisms and proteins. This becomes particularly pertinent in cases where no previous structural information is available.
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  • 文章类型: Journal Article
    电子顺磁共振(EPR)光谱与定点自旋标记(SDSL)相结合是蛋白质结构研究的有力工具。硝基氧化物是非常合适的自旋标记试剂,但是稳定性有限,特别是在细胞环境中。本文中,我们介绍了马来酰亚胺和叠氮化物修饰的四乙基保护的异吲哚啉基硝基氧(M-和Az-TEIO)的合成,用于标记半胱氨酸或非规范氨基酸对乙炔基-1-苯丙氨酸(pENF)。我们证明了TEIO位点特异性连接到蛋白质硫氧还蛋白(TRX)的高稳定性,可抵抗原核和真核环境中的还原,并进行双电子-电子共振(DEER)测量。我们进一步产生新残基PENF-Az-TEIO的旋转异构体文库,其提供与测量的分布一致的距离分布。
    Electron paramagnetic resonance (EPR) spectroscopy in combination with site-directed spin labeling (SDSL) is a powerful tool in protein structural research. Nitroxides are highly suitable spin labeling reagents, but suffer from limited stability, particularly in the cellular environment. Herein we present the synthesis of a maleimide- and an azide-modified tetraethyl-shielded isoindoline-based nitroxide (M- and Az-TEIO) for labeling of cysteines or the noncanonical amino acid para-ethynyl-l-phenylalanine (pENF). We demonstrate the high stability of TEIO site-specifically attached to the protein thioredoxin (TRX) against reduction in prokaryotic and eukaryotic environments, and conduct double electron-electron resonance (DEER) measurements. We further generate a rotamer library for the new residue pENF-Az-TEIO that affords a distance distribution that is in agreement with the measured distribution.
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