Probing the Lysine Proximal Microenvironments within Membrane Protein Complexes by Active Dimethyl Labeling and Mass Spectrometry

Ye Zhou, Yue Wu, Mingdong Yao, Zheyi Liu, Jin Chen, Jun Chen, Nobuo Kamiya Lirong Tian, Guangye Han, Jian-Ren Shen, Fangjun Wang

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Positively charged lysines are crucial to maintaining the native structures of proteins and protein complexes by forming hydrogen bonds and electrostatic interactions with their proximal amino acid residues. However, it is still a challenge to develop an efficient method for probing the active proximal microenvironments of lysines without changing their biochemical/physical properties. Herein, we developed an active covalent labeling strategy combined with mass spectrometry to systematically probe the lysine proximal microenvironments within membrane protein complexes (∼700 kDa) with high throughput. Our labeling strategy has the advantages of high labeling efficiency and stability, preservation of the active charge states, as well as biological activity of the labeled proteins. In total, 121 lysines with different labeling levels were obtained for the photosystem II complexes from cyanobacteria, red algae, and spinach and provided important insights for understanding the conserved and nonconserved local structures of PSII complexes among evolutionarily divergent species that perform photosynthesis.

Original languageEnglish
Pages (from-to)12060-12065
Number of pages6
JournalAnalytical Chemistry
Volume88
Issue number24
DOIs
Publication statusPublished - Dec 20 2016

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Labeling
Lysine
Mass spectrometry
Membrane Proteins
Proteins
Photosystem II Protein Complex
Photosynthesis
Algae
Coulomb interactions
Bioactivity
Hydrogen bonds
Physical properties
Throughput
Amino Acids

Keywords

  • Journal Article

Cite this

Probing the Lysine Proximal Microenvironments within Membrane Protein Complexes by Active Dimethyl Labeling and Mass Spectrometry. / Zhou, Ye; Wu, Yue; Yao, Mingdong; Liu, Zheyi; Chen, Jin; Chen, Jun; Tian, Nobuo Kamiya Lirong; Han, Guangye; Shen, Jian-Ren; Wang, Fangjun.

In: Analytical Chemistry, Vol. 88, No. 24, 20.12.2016, p. 12060-12065.

Research output: Contribution to journalArticle

Zhou, Ye ; Wu, Yue ; Yao, Mingdong ; Liu, Zheyi ; Chen, Jin ; Chen, Jun ; Tian, Nobuo Kamiya Lirong ; Han, Guangye ; Shen, Jian-Ren ; Wang, Fangjun. / Probing the Lysine Proximal Microenvironments within Membrane Protein Complexes by Active Dimethyl Labeling and Mass Spectrometry. In: Analytical Chemistry. 2016 ; Vol. 88, No. 24. pp. 12060-12065.
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AU - Chen, Jin

AU - Chen, Jun

AU - Tian, Nobuo Kamiya Lirong

AU - Han, Guangye

AU - Shen, Jian-Ren

AU - Wang, Fangjun

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