Real-time dynamic single-molecule protein sequencing on an integrated semiconductor device

被引：57

作者：

Reed, Brian D. ^{[1
]}

Meyer, Michael J. ^{[1
]}

Abramzon, Valentin ^{[1
]}

Ad, Omer ^{[1
]}

Adcock, Pat ^{[1
]}

Ahmad, Faisal R. ^{[1
]}

Alppay, Gun ^{[1
]}

Ball, James A. ^{[1
]}

Beach, James ^{[1
]}

Belhachemi, Dominique ^{[1
]}

Bellofiore, Anthony ^{[1
]}

Bellos, Michael ^{[1
]}

Beltran, Juan Felipe ^{[1
]}

Betts, Andrew ^{[1
]}

Bhuiya, Mohammad Wadud ^{[1
]}

Blacklock, Kristin ^{[1
]}

Boer, Robert ^{[1
]}

Boisvert, David ^{[1
]}

Brault, Norman D. ^{[1
]}

Buxbaum, Aaron ^{[1
]}

Caprio, Steve ^{[1
]}

Choi, Changhoon ^{[1
]}

Christian, Thomas D. ^{[1
]}

Clancy, Robert ^{[1
]}

Clark, Joseph ^{[1
]}

Connolly, Thomas ^{[1
]}

Croce, Kathren Fink ^{[1
]}

Cullen, Richard ^{[1
]}

Davey, Mel ^{[1
]}

Davidson, Jack ^{[1
]}

Elshenawy, Mohamed M. ^{[1
]}

Ferrigno, Michael ^{[1
]}

Frier, Daniel ^{[1
]}

Gudipati, Saketh ^{[1
]}

Hamill, Stephanie ^{[1
]}

He, Zhaoyu ^{[1
]}

Hosali, Sharath ^{[1
]}

Huang, Haidong ^{[1
]}

Huang, Le ^{[1
]}

Kabiri, Ali ^{[1
]}

Kriger, Gennadiy ^{[1
]}

Lathrop, Brittany ^{[1
]}

Li, An ^{[1
]}

Lim, Peter ^{[1
]}

Liu, Stephen ^{[1
]}

Luo, Feixiang ^{[1
]}

Lv, Caixia ^{[1
]}

Ma, Xiaoxiao ^{[1
]}

McCormack, Evan ^{[1
]}

Millham, Michele ^{[1
]}

机构：

[1] Quantum Si Inc, Guilford, CT 06437 USA

[2] Univ PSL, ESPCI Paris, CNRS UMR 8231, Lab Biochimie, Paris, France

[3] Rutgers State Univ, Dept Biochem & Mol Biol, Piscataway, NJ 08854 USA

[4] Univ Wollongong, Mol Horizons, Wollongong, NSW 2522, Australia

来源：

SCIENCE | 2022年 / 378卷 / 6616期

关键词：

STRUCTURAL BASIS; COMPLEX; CLPS;

D O I：

10.1126/science.abo7651

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页码：186 / 191

页数：6

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