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 language | English |
|---|---|
| Pages (from-to) | 12060-12065 |
| Number of pages | 6 |
| Journal | Analytical Chemistry |
| Volume | 88 |
| Issue number | 24 |
| DOIs | |
| Publication status | Published - Dec 20 2016 |
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- Journal Article
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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 journal › Article
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TY - JOUR
T1 - Probing the Lysine Proximal Microenvironments within Membrane Protein Complexes by Active Dimethyl Labeling and Mass Spectrometry
AU - Zhou, Ye
AU - Wu, Yue
AU - Yao, Mingdong
AU - Liu, Zheyi
AU - Chen, Jin
AU - Chen, Jun
AU - Tian, Nobuo Kamiya Lirong
AU - Han, Guangye
AU - Shen, Jian-Ren
AU - Wang, Fangjun
PY - 2016/12/20
Y1 - 2016/12/20
N2 - 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.
AB - 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.
KW - Journal Article
U2 - 10.1021/acs.analchem.6b02502
DO - 10.1021/acs.analchem.6b02502
M3 - Article
C2 - 28193046
VL - 88
SP - 12060
EP - 12065
JO - Analytical Chemistry
JF - Analytical Chemistry
SN - 0003-2700
IS - 24
ER -