研究人员展示了单分子蛋白质测序的动态方法，其中单肽被染料标记的N端氨基酸识别器的混合物实时探测并同时被氨基肽酶切割。研究人员通过测量集成半导体芯片上的荧光强度、寿命和结合动力学来注释氨基酸并识别肽的序列。结果证明了识别器能够以信息丰富的方式识别多个氨基酸的动力学原理，并能够辨别单一氨基酸的替换和翻译后修饰。
随着进一步的发展，研究人员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、可扩展并可实现的平台。
据介绍，蛋白质组的研究将大大受益于直接测序和数字量化蛋白质的方法，并以单分子灵敏度检测翻译后修饰。
附：英文原文
Title: Real-time dynamic single-molecule protein sequencing on an integrated semiconductor device
Author: Brian D. Reed, Michael J. Meyer, Valentin Abramzon, Omer AdPhD, Omer Ad, Pat Adcock, Faisal R. Ahmad, Gün Alppay, James A. Ball, James Beach, Dominique Belhachemi, Anthony Bellofiore, Michael Bellos, Juan Felipe Beltrán, Andrew Betts, Mohammad Wadud Bhuiya, Kristin Blacklock, Robert Boer, David Boisvert, Norman D. Brault, Aaron Buxbaum, Steve Caprio, Changhoon Choi, Thomas D. Christian, Robert Clancy, Joseph Clark, Thomas Connolly, Kathren Fink Croce, Richard Cullen, Mel Davey, Jack Davidson, Mohamed M. Elshenawy, Michael Ferrigno, Daniel Frier, Saketh Gudipati, Stephanie Hamill, Zhaoyu He, Sharath Hosali, Haidong Huang, Le Huang, Ali Kabiri, Gennadiy Kriger, Brittany Lathrop, An Li, Peter Lim, Stephen Liu, Feixiang Luo, Caixia Lv, Xiaoxiao Ma, Evan McCormack, Michele Millham, Roger Nani, Manjula Pandey, John Parillo, Gayatri Patel, Douglas H. Pike, Kyle Preston, Adeline Pichard-Kostuch, Kyle Rearick, Todd Rearick, Marco Ribezzi-Crivellari, Gerard Schmid, Jonathan Schultz, Xinghua Shi, Badri Singh, Nikita Srivastava, Shannon F. Stewman, TR Thurston, T. R. Thurston, Philip Trioli, Jennifer Tullman, Xin Wang, Yen-Chih Wang, Eric A. G. Webster, Zhizhuo Zhang, Jorge Zuniga, Smita S. Patel, Andrew D. Griffiths, Antoine M. van Oijen, Michael McKenna, Matthew D. Dyer, Jonathan M. Rothberg
Issue&Volume: 2022-10-14
Abstract: 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.
DOI: abo7651
Source: https://www.science.org/doi/10.1126/science.abo7651