Abstract Realization of binding reagents that can perform high-fidelity, sequential recognition and detection of amino acids is important for the success of many proposed approaches for single-molecule protein sequencing. Towards this purpose, a variant of the Agrobacterium tumefaciens protein ClpS was previously engineered with improved binding affinity for phenylalanine at the N-terminus of a peptide. In this study, this variant was further engineered for attributes necessary for robust application as a biotechnology reagent such as increased thermostability and improved selectivity under different buffer conditions. The stabilized variant was further characterized in terms of selectivity for the N-terminal residue in the context of different peptides with varying residues at positions 2 (P2) and 3 (P3). The implication of the study for the use of ClpS based reagents in peptide sequencing technologies are described.

中文翻译：

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基于 ClpS 的 N 端氨基酸结合试剂，具有更高的热稳定性和选择性

摘要 实现对氨基酸进行高保真、顺序识别和检测的结合试剂对于许多提议的单分子蛋白质测序方法的成功至关重要。为了这个目的，根癌农杆菌蛋白 ClpS 的一种变体先前被设计为具有改进的对肽 N 端苯丙氨酸的结合亲和力。在这项研究中，该变体经过进一步设计，具有作为生物技术试剂稳健应用所必需的属性，例如在不同缓冲条件下提高热稳定性和提高选择性。在位置2(P2)和3(P3)具有不同残基的不同肽的背景下，根据对N-末端残基的选择性进一步表征稳定化变体。