Hyperpolarised NMR to follow water proton transport through membrane channels via exchange with biomolecules

被引:6
|
作者
Nastasa, Viorel [1 ,2 ]
Stavarache, Cristina [3 ,4 ]
Hanganu, Anamaria [3 ,4 ]
Coroaba, Adina [5 ]
Nicolescu, Alina [4 ,5 ]
Deleanu, Calin [4 ,5 ]
Sadet, Aude [3 ]
Vasos, Paul R. [1 ,3 ]
机构
[1] Horia Hulubei Inst Nucl Phys IFIN HH, ELI NP, Reactorului Str 30,Magurele Campus, Bucharest, Romania
[2] Natl Inst Laser Plasma & Radiat Phys, Atomistilor Str 409, Magurele, Romania
[3] Univ Bucharest ICUB, Res Inst, 36-46 Blvd M Kogalniceanu, RO-050107 Bucharest, Romania
[4] CD Nenitescu Ctr Organ Chem, 202-B Spl Independentei, RO-060023 Bucharest, Romania
[5] Petru Poni Inst Macromol Chem, 41-A Aleea Grigore Ghica Voda, RO-700487 Iasi, Romania
基金
欧盟地平线“2020”;
关键词
NUCLEAR-MAGNETIC-RESONANCE; LONG-LIVED STATES; FATTY-ACIDS; PERMEABILITY COEFFICIENT; MOLECULAR-TRANSPORT; CARBOXYLIC-ACIDS; LIPID-BILAYERS; PERMEATION; KINETICS; SPECTROSCOPY;
D O I
10.1039/c8fd00021b
中图分类号
O64 [物理化学(理论化学)、化学物理学];
学科分类号
070304 ; 081704 ;
摘要
Water uptake in vesicles and the subsequent exchange between water protons and amide -NH protons in amino acids can be followed by a new, highly sensitive, type of magnetic resonance spectroscopy: dynamic nuclear polarisation (DNP)-enhanced NMR in the liquid state. Water hydrogen atoms are detected prior to and after their transfer to molecular sites in peptides and proteins featuring highly-accessible proton-exchangeable groups, as is the case for the -NH groups of intrinsically disordered proteins. The detected rates for amide proton-water proton exchange can be modulated by membrane-crossing rates, when a membrane channel is interposed. We hyperpolarised water proton spins via dynamic nuclear polarisation followed by sample dissolution (d-DNP) and transferred the created polarisation to -NH groups with high solvent accessibility in an intrinsically disordered protein domain. This domain is the membrane anchor of c-Src kinase, whose activity controls cell proliferation. The hindrance of effective water proton transfer rate constants observed in free solvent when a membrane-crossing step is involved is discussed. This study aims to assess the feasibility of recently-introduced hyperpolarised (DNP-enhanced) NMR to assess water membrane crossing dynamics.
引用
收藏
页码:67 / 82
页数:16
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