Whole cell solid-state NMR study of Chlamydomonas reinhardtii microalgae

被引:34
作者
Arnold, Alexandre A. [1 ]
Bourgouin, Jean-Philippe [1 ]
Genard, Bertrand [1 ,2 ]
Warschawski, Dror E. [1 ,3 ,4 ]
Tremblay, Rejean [2 ]
Marcotte, Isabelle [1 ]
机构
[1] Univ Quebec Montreal, Dept Chem, Downtown Stn, POB 8888, Montreal, PQ H3C 3P8, Canada
[2] Univ Quebec Rimouski, Inst Sci Mer Rimouski, 310 Allee Ursulines, Rimouski, PQ G5L 3A1, Canada
[3] Univ Paris Diderot, CNRS, UMR 7099, Lab Biol Physicochim Prot Membranaires, 13 Rue Pierre & Marie Curie, F-75005 Paris, France
[4] IBPC, 13 Rue Pierre & Marie Curie, F-75005 Paris, France
基金
加拿大自然科学与工程研究理事会; 加拿大健康研究院;
关键词
In vivo NMR; Magic-angle spinning; Lipids; Cell-wall; Starch; Isotope labelling; Dynamic filters; C-13; CHEMICAL-SHIFTS; CHLOROPLAST MEMBRANES; MAS NMR; POLYSACCHARIDES; IDENTIFICATION; EVOLUTION; DYNAMICS; PROTEIN;
D O I
10.1007/s10858-018-0164-7
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
In vivo or whole-cell solid-state NMR is an emerging field which faces tremendous challenges. In most cases, cell biochemistry does not allow the labelling of specific molecules and an in vivo study is thus hindered by the inherent difficulty of identifying, among a formidable number of resonances, those arising from a given molecule. In this work we examined the possibility of studying, by solid-state NMR, the model organism Chlamydomonas reinhardtii fully and non-specifically C-13 labelled. The extension of NMR-based dynamic filtering from one-dimensional to two-dimensional experiments enabled an enhanced selectivity which facilitated the assignment of cell constituents. The number of resonances detected with these robust and broadly applicable experiments appears to be surprisingly sparse. Various constituents, notably galactolipids abundant in organelle membranes, carbohydrates from the cell wall, and starch from storage grains could be unambiguously assigned. Moreover, the dominant crystal form of starch could be determined in situ. This work illustrates the feasibility and caveats of using solid-state NMR to study intact non-specifically C-13 labelled micro-organisms.
引用
收藏
页码:123 / 131
页数:9
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