Analysis of enzymatically generated phosphoinositides by 31P nuclear magnetic resonance spectroscopy

Müller, Mat­thias; Schiller, Jürgen; Petković, Marijana; Zschörnig, Olaf; Arnhold, Jürgen; Arnold, Klaus

doi:10.1016/j.ab.2004.03.071

Cited by 23 publications

(3 citation statements)

References 19 publications

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“…31 P MAS ssNMR revealed significant chemical shift perturbations as a function of pH (Figure S7). Based on (Kishore and Prestegard, 2003; Müller et al, 2004), we tentatively assigned the four visible signals, from higher to lower chemical shifts: 4-phosphate (4-P)-, 4,5-bisphosphate (4,5-P 2 )-monoester, PS and PC. The diester groups might be obscured by the PS and PC signal.…”

Section: Resultsmentioning

confidence: 98%

Nanodomain Clustering of the Plant Protein Remorin by Solid-State NMR

Legrand

Martinez

Grélard

et al. 2019

Front. Mol. Biosci.

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Nanodomains are dynamic membrane subcompartments, enriched in specific lipid, and protein components that act as functional platforms to manage an abundance of cellular processes. The remorin protein of plants is a well-established nanodomain marker and widely serves as a paradigm to study nanodomain clustering. Located at the inner leaflet of the plasma membrane, remorins perform essential functions during signaling. Using deuterium and phosphorus solid-state NMR, we inquire on the molecular determinants of the lipid-protein and protein-protein interactions driving nanodomain clustering. By monitoring thermotropism properties, lipid acyl chain order and membrane thickness, we report the effects of phosphoinositides and sterols on the interaction of various remorin peptides and protein constructs with the membrane. We probed several critical residues involved in this interaction and the involvement of the coiled-coil homo-oligomerisation domain into the formation of remorin nanodomains. We trace the essential role of the pH in nanodomain clustering based on anionic lipids such as phosphoinositides. Our results reveal a complex interplay between specific remorin residues and domains, the environmental pH and their resulting effects on the lipid dynamics for phosphoinositide-enriched membranes.

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Section: Resultsmentioning

confidence: 98%

Nanodomain Clustering of the Plant Protein Remorin by Solid-State NMR

Legrand

Martinez

Grélard

et al. 2019

Front. Mol. Biosci.

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“…This is due to the fact that in the absence of detergents, PLs spontaneously form vesicles in water, resulting in relatively immobile PLs. Thus, vesicle-to-micelle transformation is essential to PL solubilization by detergents (Lopez et al 1998;Muller et al 2004;Puppato et al 2007;Schiller et al 2001;Pearce and Komoroski 1993;Fuchs et al 2007a, b;Sullentrop et al 2002;Jakop et al 2009;Renner et al 2005;Dannenberger et al 2010;Bauer et al 2009). A number of arguments speak in favor of PL solubilization by detergents, e.g., the possibility to prepare samples by directly dissolving membranes or biological tissue without the need for extraction, and to achieve narrow LWs even at high PL concentrations (London and Feigenson 1979).…”

Section: Detergent-containing Solventsmentioning

confidence: 99%

Principles of multiparametric optimization for phospholipidomics by 31P NMR spectroscopy

Lutz

Cozzone

2013

Biophys Rev

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Phospholipids have long been known to be the principal constituents of the bilayer matrix of cell membranes. While the main function of cell membranes is to provide physical separation between intracellular and extracellular compartments, further biological and biochemical functions for phospholipids have been identified more recently, notably in cell signaling, cell recognition and cell-cell interaction, but also in cell growth, electrical insulation of neurons and many other processes. Therefore, accurate and efficient determination of tissue phospholipid composition is essential for our understanding of biological tissue function. P NMR spectroscopy is a quantitative and fast method for analyzing phospholipid extracts from biological samples without prior separation. However, the number of phospholipid classes and subclasses that can be quantified separately and reliably inP NMR spectra of tissue extracts is critically dependent on a variety of experimental conditions. Until recently, little attention has been paid to the optimization of phospholipid P NMR spectra. This review surveys the basic physicochemical properties that determine the quality of phospholipid spectra, and describes an optimization strategy based on this assessment. Notably, the following experimental parameters need to be controlled for systematic optimization: (1) extract concentration, (2) concentration of chelating agent, (3) pH value of the aqueous component of the solvent system, and (4) temperature of the NMR measurement. We conclude that a multiparametric optimization approach is crucial to obtaining highly predictable and reproducibleP NMR spectra of phospholipids.

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“…As the PLs in these mixed micelles are not clumped tightly to each other it is even possible to resolve resonances corresponding to PLs of the same class but with different acyl chain composition (Pearce et al, 1993;Pearce and Komoroski, 2000;Komoroski et al, 2001;Schiller and Arnold, 2002). Because the spectral region in which most of the PL-resonances appear is relatively small, only extending from −1 to 5 ppm (Gorestein, 1984;Muller et al, 2004), and due to the sensitivity of some chemical shifts to temperature and pH, possible spectral overlaps and variations in chemical shift can lead to incorrect assignments and erroneous quantification of PL composition. To address these concerns, this study was carried out to (a) enhance the accuracy in the assignment of 31 P NMR spectral resonances by determining the temperature and pH-dependence of several common PLs, and (b) to determine experimental conditions that reduce spectral overlaps and could, therefore, minimize errors in the quantitative analysis of PLs.…”

Section: Introductionmentioning

confidence: 99%