Interaction of membrane proteins and lipids with solubilizing detergents

Maire, Marc le; Champeil, Philippe; Møller, Jesper

doi:10.1016/s0304-4157(00)00010-1

Cited by 910 publications

(795 citation statements)

References 198 publications

Supporting

Mentioning

734

Contrasting

Unclassified

Order By: Relevance

“…The development of a rational approach to the solubilization of membrane proteins always requires a review of the literature to determine the detergent type and the conditions that are generally used for the solubilization of the protein of interest (1,(3)(4)(5)(6)(7)(8)10). All assays were carried at 37ºC as described in Methods.…”

Section: Discussionmentioning

confidence: 99%

“…Several excellent reviews can be found on detergents, their physical properties and their use in the solubilization of membrane proteins (1)(2)(3)(4). Due to three important factors, non-ionic detergents are frequently used for the solubilization of protein membranes: i) their efficiency in breaking the lipid-lipid and lipid-protein interactions, ii) their inefficiency in weakening protein-protein interactions, and iii) their property of being less denaturing than the ionic detergents (3)(4)(5)(6).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Solubilization of Na,K-ATPase from rabbit kidney outer medulla using only C12E8

Santos

Lamas

Ciancaglini

2002

Braz J Med Biol Res

View full text Add to dashboard Cite

SDS, C 12 E 8 , CHAPS or CHAPSO or a combination of two of these detergents is generally used for the solubilization of Na,K-ATPase and other ATPases. Our method using only C 12 E 8 has the advantage of considerable reduction of the time for enzyme purification, with rapid solubilization and purification in a single chromatographic step. Na,KATPase-rich membrane fragments of rabbit kidney outer medulla were obtained without adding SDS. Optimum conditions for solubilization were obtained at 4ºC after rapid mixing of 1 mg of membrane Na,K-ATPase with 1 mg of C 12 E 8 /ml, yielding 98% recovery of the activity. The solubilized enzyme was purified by gel filtration on a Sepharose 6B column at 4ºC. Non-denaturing PAGE revealed a single protein band with phosphomonohydrolase activity. The molecular mass of the purified enzyme estimated by gel filtration chromatography was 320 kDa. The optimum apparent pH obtained for the purified enzyme was 7.5 for both PNPP and ATP. The dependence of ATPase activity on ATP concentration showed high (K 0.5 = 4.0 µM) and low (K 0.5 = 1.4 mM) affinity sites for ATP, with negative cooperativity. Ouabain (5 mM), oligomycin (1 µg/ml) and sodium vanadate (3 µM) inhibited the ATPase activity of C 12 E 8 -solubilized and purified Na,KATPase by 99, 81 and 98.5%, respectively. We have shown that Na,KATPase solubilized only with C 12 E 8 can be purified and retains its activity. The activity is consistent with the form of (aß) 2 association.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Solubilization of Na,K-ATPase from rabbit kidney outer medulla using only C12E8

Santos

Lamas

Ciancaglini

2002

Braz J Med Biol Res

View full text Add to dashboard Cite

show abstract

“…However, this trajectory is just a suggestion, as it is difficult to follow each peak unambiguously. (33,34,51)). Line broadening can be seen at intermediate detergent concentrations (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Our study focuses on TRP3 accommodation at the hydrophilic/ hydrophobic interface of micelles and subsequent events. The structure and dynamics of TRP3 were examined in the presence of micelles of different chemical nature: zwitterionic (lysolauroylphosphatidylcholine 1-lauroyl-2-hydroxy-sn-glycero-3-phosphocholine (LLPC), n-dodecylphosphocholine (DPC), and N-dodecyl-N,N-(dimethylammonio) butyrate (DDMAB), anionic 1-myristoyl-2-hydroxy-sn-glycero-3-phospho-(1 0 -rac-glycerol) (LMPG), and nonionic (octylglucoside (OG)) (23,24,(31)(32)(33)(34)(35). The conformational ensemble was evaluated during detergent titration and it was found that the peptide interacts via conformational selection.…”

Section: Introductionmentioning

confidence: 99%

Structural and Dynamic Insights of the Interaction between Tritrpticin and Micelles: An NMR Study

Santos

Moraes

Nakaie

et al. 2016

Biophysical Journal

View full text Add to dashboard Cite

A large number of antimicrobial peptides (AMPs) acts with high selectivity and specificity through interactions with membrane lipid components. These peptides undergo complex conformational changes in solution; upon binding to an interface, one major conformation is stabilized. Here we describe a study of the interaction between tritrpticin (TRP3), a cathelicidin AMP, and micelles of different chemical composition. The peptide's structure and dynamics were examined using one-dimensional and two-dimensional NMR. Our data showed that the interaction occurred by conformational selection and the peptide acquired similar structures in all systems studied, despite differences in detergent headgroup charge or dipole orientation. Fluorescence and paramagnetic relaxation enhancement experiments showed that the peptide is located in the interface region and is slightly more deeply inserted in 1-myristoyl-2-hydroxy-sn-glycero-3-phospho-1 0 -rac-glycerol (LMPG, anionic) than in 1-lauroyl-2-hydroxy-sn-glycero-3-phosphocholine (LLPC, zwitterionic) micelles. Moreover, the tilt angle of an assumed helical portion of the peptide is similar in both systems. In previous work we proposed that TRP3 acts by a toroidal pore mechanism. In view of the high hydrophobic core exposure, hydration, and curvature presented by micelles, the conformation of TRP3 in these systems could be related to the peptide's conformation in the toroidal pore.

show abstract

“…As can be seen in Figure 2, the active surfaces of the labwares become saturated by the CHAPS monomers, and once the sample is introduced in the sampling bottle, the analytes are available for extraction. Additionally, it was observed that for 12 complete elimination of adsorption losses from the 96-well plate, a concentration above the critical micelle concentration (CMC, 3-10 mM 14 ) is important, particularly for the most non polar compounds in the study. This also shows that CHAPS is not only saturating the lab-ware surfaces, it is also able to stabilize the analytes in the micelles/sample (Figure 2).…”

Section: Theoretical Consideration Of Micellar Spme and Micelle Assismentioning

confidence: 99%

Micelle Assisted Thin-Film Solid Phase Microextraction: A New Approach for Determination of Quaternary Ammonium Compounds in Environmental Samples

Boyacı

Pawliszyn

2014

Anal. Chem.

View full text Add to dashboard Cite

Keywords: High throughput analysis, Quaternary ammonium compounds, Sample preparation of challenging compounds, Thin-film solid phase microextraction, Micelle-modified matrix, micelleassisted stabilization of quaternary ammonium compounds, micelle assisted TF-SPME 2 ABSTRACTDetermination of quaternary ammonium compounds (QACs) often is considered to be a challenging undertaking owing to secondary interactions of the analytes' permanently charged quaternary ammonium head or hydrophobic tail with the utilized lab-wares.Here, for the first time, a micelle assisted thin-film solid phase microextraction (TF-SPME) using a zwitterionic detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) as a matrix modifier is introduced as a novel approach for in-laboratory sample preparation of the challenging compounds.The proposed micelle assisted thin-film solid phase microextraction (TF-SPME) method offers suppression/enhancement free electrospray ionization of analytes in mass spectrometric detection, minimal interaction of the micelles with the TF-SPME coating, and chromatographic stationary phase and analysis free of secondary interactions. Moreover, it was found that the matrix modifier has multiple functions; when its concentration is found below the critical micelle concentration (CMC), the matrix modifier primarily act as a surface deactivator; above its CMC, it acts as a stabilizer for QACs. Additionally, shorter equilibrium extraction times in the presence of the modifier demonstrated that micelles also assist in the transfer of analytes from the bulk of the sample to the surface of the coating.The developed micelle assisted TF-SPME protocol using the 96-blade system requires only 30 min of extraction and 15 min of desorption. Together with a conditioning step (15 min), the entire method is 60 min; considering the advantage of using the 96-blade system, if all the blades in the brush are used, the sample preparation time per sample is 0.63 min. Moreover, the recoveries for all analytes with the developed method were found to range within 80.2%-97.3%; as such, this 3 method can be considered an open bed solid phase extraction. The proposed method was successfully validated using real samples.4

show abstract

Interaction of membrane proteins and lipids with solubilizing detergents

Cited by 910 publications

References 198 publications

Solubilization of Na,K-ATPase from rabbit kidney outer medulla using only C12E8

Solubilization of Na,K-ATPase from rabbit kidney outer medulla using only C12E8

Structural and Dynamic Insights of the Interaction between Tritrpticin and Micelles: An NMR Study

Micelle Assisted Thin-Film Solid Phase Microextraction: A New Approach for Determination of Quaternary Ammonium Compounds in Environmental Samples

Contact Info

Product

Resources

About