Leveraging nature’s biomolecular designs in next-generation protein sequencing reagent development
Leveraging nature’s biomolecular designs in next-generation protein sequencing reagent development
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DOI：
10.1007/s00253-020-10745-2
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发表时间：
2020-07
影响因子：
5
通讯作者：
J. Tullman;J. Marino;Z. Kelman
中科院分区：
文献类型：
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作者：
J. Tullman;J. Marino;Z. Kelman
下一代蛋白质测序方法正在兴起，有可能彻底改变蛋白质组学领域。一种这样的测序方法涉及基于荧光的固定化肽成像，其中通过一系列荧光标记的生物分子顺序读出多肽的N端氨基酸。当选择性结合特定的 N 端氨基酸时，NAAB（N 端氨基酸结合剂）亲和试剂通过其相关的荧光标签识别该氨基酸。实施这种荧光测序方法的一个关键技术挑战是需要开发对这种生物技术应用所需的特定 N 末端氨基酸具有高亲和力和选择性的 NAAB 亲和试剂。开发此类 NAAB 亲和试剂的一种方法是利用结合氨基酸和/或肽的天然生物分子。在这里，我们描述了可以考虑用于此目的的几种候选生物分子，并讨论了每种生物分子的开发潜力。关键点•下一代测序方法的出现可能会彻底改变蛋白质组学。•通过荧光标记的亲和试剂连续读出N端氨基酸。•天然肽/氨基酸结合物可以设计成亲和试剂。•蛋白质的大小和结构有助于试剂开发的可行性。
Next-generation approaches for protein sequencing are now emerging that could have the potential to revolutionize the field in proteomics. One such sequencing method involves fluorescence-based imaging of immobilized peptides in which the N-terminal amino acid of a polypeptide is readout sequentially by a series of fluorescently labeled biomolecules. When selectively bound to a specific N-terminal amino acid, the NAAB (N-terminal amino acid binder) affinity reagent identifies the amino acid through its associated fluorescence tag. A key technical challenge in implementing this fluoro-sequencing approach is the need to develop NAAB affinity reagents with the high affinity and selectivity for specific N-terminal amino acids required for this biotechnology application. One approach to develop such a NAAB affinity reagent is to leverage naturally occurring biomolecules that bind amino acids and/or peptides. Here, we describe several candidate biomolecules that could be considered for this purpose and discuss the potential for developability of each.Key points• Next-generation sequencing methods are emerging that could revolutionize proteomics.• Sequential readout of N-terminal amino acids by fluorescent-tagged affinity reagents.• Native peptide/amino acid binders can be engineered into affinity reagents.• Protein size and structure contribute to feasibility of reagent developability.