title: Probing the Lysine Proximal Microenvironments within Membrane Protein Complexes by Active Dimethyl Labeling and Mass Spectrometry.
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.
authors.affiliation: Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , Dalian 116023, China.|University of Chinese Academy of Sciences , Beijing 100049, China.
authors.forename: Ye
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , Dalian 116023, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Wu
authors.affiliation: State Key Laboratory of Catalysis, Dalian National Laboratory for Clean Energy, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , Dalian 116023, China.
authors.forename: Mingdong
authors.initials: M
authors.lastname: Yao
authors.affiliation: Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , Dalian 116023, China.|University of Chinese Academy of Sciences , Beijing 100049, China.
authors.forename: Zheyi
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , Dalian 116023, China.|University of Chinese Academy of Sciences , Beijing 100049, China.
authors.forename: Jin
authors.initials: J
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Catalysis, Dalian National Laboratory for Clean Energy, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , Dalian 116023, China.
authors.forename: Jun
authors.identifier: 0000-0002-4114-3046
authors.initials: J
authors.lastname: Chen
authors.affiliation: Key Laboratory of Photobiology, Institute of Botany, The Chinese Academy of Sciences , Beijing 100093, China.|University of Chinese Academy of Sciences , Beijing 100049, China.
authors.forename: Lirong
authors.initials: L
authors.lastname: Tian
authors.affiliation: Key Laboratory of Photobiology, Institute of Botany, The Chinese Academy of Sciences , Beijing 100093, China.
authors.forename: Guangye
authors.initials: G
authors.lastname: Han
authors.affiliation: Key Laboratory of Photobiology, Institute of Botany, The Chinese Academy of Sciences , Beijing 100093, China.|Photosynthesis Research Center, Graduate School of Natural Science and Technology, Okayama University , 1-1, Naka 3-chome, Tsushima, Okayama 700-8530, Japan.
authors.forename: Jian-Ren
authors.initials: JR
authors.lastname: Shen
authors.affiliation: Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , Dalian 116023, China.
authors.forename: Fangjun
authors.identifier: 0000-0002-8118-7019
authors.initials: F
authors.lastname: Wang
mesh_terms: D001894:Borohydrides / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D018360:Crystallography, X-Ray; D000458:Cyanobacteria / Q000378:metabolism; D005557:Formaldehyde / Q000737:chemistry; D008239:Lysine / Q000737:chemistry*; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism; D009211:Myoglobin / Q000737:chemistry / Q000378:metabolism; D045332:Photosystem II Protein Complex / Q000737:chemistry; D017434:Protein Structure, Tertiary; D000461:Rhodophyta / Q000378:metabolism; D018724:Spinacia oleracea / Q000378:metabolism; D053719:Tandem Mass Spectrometry
chemical_list: D001894:Borohydrides; D008565:Membrane Proteins; D009211:Myoglobin; D045332:Photosystem II Protein Complex; D005557:Formaldehyde; C009282:sodium cyanoborohydride; D008239:Lysine