Quantum-Si, Inc., Guilford, CT 06437, USA.
Laboratoire de Biochimie, ESPCI Paris, Université PSL, CNRS UMR 8231, Paris, France.
Science. 2022 Oct 14;378(6616):186-192. doi: 10.1126/science.abo7651. Epub 2022 Oct 13.
Studies of the proteome would benefit greatly from methods to directly sequence and digitally quantify proteins and detect posttranslational modifications with single-molecule sensitivity. Here, we demonstrate single-molecule protein sequencing using a dynamic approach in which single peptides are probed in real time by a mixture of dye-labeled N-terminal amino acid recognizers and simultaneously cleaved by aminopeptidases. We annotate amino acids and identify the peptide sequence by measuring fluorescence intensity, lifetime, and binding kinetics on an integrated semiconductor chip. Our results demonstrate the kinetic principles that allow recognizers to identify multiple amino acids in an information-rich manner that enables discrimination of single amino acid substitutions and posttranslational modifications. With further development, we anticipate that this approach will offer a sensitive, scalable, and accessible platform for single-molecule proteomic studies and applications.
蛋白质组学的研究将极大地受益于能够直接对蛋白质进行测序和数字定量，并具有单分子灵敏度来检测翻译后修饰的方法。在这里，我们展示了一种动态方法的单分子蛋白质测序，其中通过用染料标记的 N 端氨基酸识别器的混合物实时探测单个肽，并同时由氨肽酶切割。我们通过在集成半导体芯片上测量荧光强度、寿命和结合动力学来注释氨基酸并识别肽序列。我们的结果证明了允许识别器以信息丰富的方式识别多种氨基酸的动力学原理，从而能够区分单个氨基酸取代和翻译后修饰。随着进一步的发展，我们预计这种方法将为单分子蛋白质组学研究和应用提供一个敏感、可扩展和易于使用的平台。
