Online Table I parameters of the fluorescence emission spectra of mTM4 and mTM10 in various media

Online Table I Parameters of the fluorescence emission spectra of mTM4 and mTM10 in various media Peptide Medium λmax (nm) mTM4 Pure TFE 340 mTM4 TFE/H2O (50/50, v/v) 342.5 mTM4 DPC micelles 320 mTM4 DDM micelles 318.5 mTM4 Buffer 328.5 mTM10 Pure TFE 307.5 mTM10 TFE/H2O (50/50, v/v) 339 mTM10 DPC micelles 331 mTM10 DDM micelles 327 mTM10 Buffer 329.5 Standard conditions: λex=280 nm, slits 1.25 mm at excitation and emission (λmax for NATA in water was 352 nm in these conditions); 10 µM peptide; buffer: 10 mM potassium phosphate pH 7.5. Detergent was 4 mM in buffer. T=20°C Transverse and tangential orientation of predicted transmembrane fragments 4 and 10 from the human multidrug resistance protein (hMRP1/ABCC1) in membrane mimics European Biophysics Journal Béatrice de Forestaa,b Michel Vincent, Manuel Garrigos and Jacques Gallayc,d aCEA, iBiTecS, Service de Bioénergétique Biologie Structurale et Mécanismes, F-91191, Gif-sur-Yvette, France; bCNRS, URA 2096, F-91191, Gif-sur-Yvette, France; cUniversité Paris-Sud 11, UMR 8619, Institut de Biochimie et Biophysique Moléculaire et Cellulaire, Bât. 430, Orsay, France; dCNRS, UMR 8619, Orsay, France Corresponding authors. B. de Foresta: beatrice.de-foresta@cea.fr J. Gallay: jacques.gallay@u-psud.fr Online Table II Characteristics for predicted transmembrane fragments of hMRP1 Domain Peptide name Sequence Charge ΔG Unstructured (kcal/mol) MSD0 TM1 CF-QNTVLVWVPCFYLWACFPFYF-LY 0 -14.7 TM2 TK-TALGFLLWIVCWADLFYSFWE-RS ++,-- -9.21 TM3 RGY-FLAPVFLVSPTLLGITTLLAT-F + -7.02 TM4 VQ-SSGIMLTFWLVALVCALAILR-SK ++ -5.78 TM5 RD-ITFYVYFSLLLIQLVLSCFSD-RS ++,-- -6.02 MSD1 TM6 KV-LYKTFGPYFLMSFFFKAIHDL-MM 4+,- -6.63 TM7 (W)QG-YFYTVLLFVTACLQTLVLHQY-FH ++ -7.33 TM8 TY-INMIWSAPLQVILALYLLWLN-LG 0 -10.21 TM9 GP-SVLAGVAVMVLMVPVNAVMAM-KT + -0.82 TM10 SA-VGTFTWVCTPFLVALCTFAVY-VT 0 -8.18 TM11 FN-ILRFPLNILPMVISSIVQASV-SL + -4.49 MSD2 TM12 (WD)YM-KAIGLFISFLSIFLFMCNHVS-AL ++ -8.7 TM13 IS-QGIAVFGYSMAVSIGGILASR-CL + -4.85 TM14 PE-VIKMFMGSLFNVIGACIVILL-AT +,- -3.71 TM15 LA-TPIAAIIIPPLGLIYFFVQRF-YV + -7.06 TM16 ANRWLAVRL-ECVGNCIVLFAALFAV(ISRHS-LS) ++,- -4.07 TM17 (LS-)AGVGLSVSYSLQVTTTYLNWL-VRMS + -6.23 The sequences were the 21 amino-acid sequences predicted for human MRP1 (from Swissprot-Uniprot-hMRP, code P33527), to which we added the N-terminal and C-terminal amino acids from the same sequence, unless otherwise stated. TM16 and 17 were slightly more extended on the cytoplasmic side. In parentheses: Trp close to TM or Uniprot-predicted TM sequences. Hydropathy color code: Hydrophobic Polar Basic Acidic. Free energy: From MPEX 3.0, N-T acetylated, C-T CONH2, partitioning from water to bilayer, TM: n=25 Transverse and tangential orientation of predicted transmembrane fragments 4 and 10 from the human multidrug resistance protein (hMRP1/ABCC1) in membrane mimics European Biophysics Journal Béatrice de Forestaa,b Michel Vincent, Manuel Garrigos and Jacques Gallayc,d aCEA, iBiTecS, Service de Bioénergétique Biologie Structurale et Mécanismes, F-91191, Gif-sur-Yvette, France; bCNRS, URA 2096, F-91191, Gif-sur-Yvette, France; cUniversité Paris-Sud 11, UMR 8619, Institut de Biochimie et Biophysique Moléculaire et Cellulaire, Bât. 430, Orsay, France; dCNRS, UMR 8619, Orsay, France Corresponding authors. B. de Foresta: beatrice.de-foresta@cea.fr J. Gallay: jacques.gallay@u-psud.fr Online Figure I Kinetics of the titration of mTM4 with DPC and DDM We added 10 µM mTM4 to 10 mM potassium phosphate buffer, pH 7.5, at 20°C (first arrow). Aliquots of DPC (black trace) or DDM (blue trace) (series of black and blue arrows respectively) were then added sequentially, with continuous stirring, at constant intervals (~50 s). For DPC (cmc=1.1 mM), final concentrations in the cuvette were 0.5, 1, 2, 4 and 6 mM. For DDM (cmc=0.17 mM), final concentrations were 0.1, 0.2, 1, 2, 3 and 6 mM. Fluorescence intensity was recorded continuously with ex set at 280 nm and λem at 320 nm. Slit widths were 1.25 mm (bandwidths ~5 nm) for both excitation and emission Transverse and tangential orientation of predicted transmembrane fragments 4 and 10 from the human multidrug resistance protein (hMRP1/ABCC1) in membrane mimics European Biophysics Journal Béatrice de Forestaa,b Michel Vincent, Manuel Garrigos and Jacques Gallayc,d aCEA, iBiTecS, Service de Bioénergétique Biologie Structurale et Mécanismes, F-91191, Gif-sur-Yvette, France; bCNRS, URA 2096, F-91191, Gif-sur-Yvette, France; cUniversité Paris-Sud 11, UMR 8619, Institut de Biochimie et Biophysique Moléculaire et Cellulaire, Bât. 430, Orsay, France; dCNRS, UMR 8619, Orsay, France Corresponding authors. B. de Foresta: beatrice.de-foresta@cea.fr J. Gallay: jacques.gallay@u-psud.fr Online Figure II Fluorescence emission spectra of the hMRP1 fragments in various media A: Emission spectra for 10 µM mTM4 in TFE/water (50/50, v/v) (long dashed lines) or TFE (short dashed line). ex was set at 280 nm. Slit widths were 1.25 mm (bandwidths ~5 nm) for both excitation and emission. Spectra were recorded after a short period of equilibration (2-3 min) and the readings for background spectra (same solvent or detergent in buffer) were subtracted. B: Emission spectra for 10 µm mTM10. Same symbols as in panel A. In both panels, the emission spectrum of 10 µM NATA in pure water is indicated as a reference (green line) Transverse and tangential orientation of predicted transmembrane fragments 4 and 10 from the human multidrug resistance protein (hMRP1/ABCC1) in membrane mimics European Biophysics Journal Béatrice de Forestaa,b Michel Vincent, Manuel Garrigos and Jacques Gallayc,d aCEA, iBiTecS, Service de Bioénergétique Biologie Structurale et Mécanismes, F-91191, Gif-sur-Yvette, France; bCNRS, URA 2096, F-91191, Gif-sur-Yvette, France; cUniversité Paris-Sud 11, UMR 8619, Institut de Biochimie et Biophysique Moléculaire et Cellulaire, Bât. 430, Orsay, France; dCNRS, UMR 8619, Orsay, France Corresponding authors. B. de Foresta: beatrice.de-foresta@cea.fr J. Gallay: jacques.gallay@u-psud.fr Online Figure III Fluorescence emission spectra of mTM4 in mixed micelles of BrDDM/DDM (panel A) or BrUM/DDM (panel B) A: Emission spectra for 5 µM mTM4 in 10 mM potassium phosphate buffer, pH 7.5, supplemented with various mixtures of BrDDM and DDM, at a final total detergent concentration of 4 mM, at 20°C. X, the molar fraction of BrDDM was 0, 0.1, 0.2, 0.4, 0.6 or 1 (from upper to lower spectra). ex was set at 280 nm. Slit widths were 1.25 mm (bandwidths ~5 nm) for both excitation and emission. Spectra were recorded after a short period of equilibration (~3 min) and the readings for background spectra (detergent in buffer) were subtracted. B: Similar experiment to that in panel A, with BrUM as the brominated detergent Transverse and tangential orientation of predicted transmembrane fragments 4 and 10 from the human multidrug resistance protein (hMRP1/ABCC1) in membrane mimics European Biophysics Journal Béatrice de Forestaa,b Michel Vincent, Manuel Garrigos and Jacques Gallayc,d aCEA, iBiTecS, Service de Bioénergétique Biologie Structurale et Mécanismes, F-91191, Gif-sur-Yvette, France; bCNRS, URA 2096, F-91191, Gif-sur-Yvette, France; cUniversité Paris-Sud 11, UMR 8619, Institut de Biochimie et Biophysique Moléculaire et Cellulaire, Bât. 430, Orsay, France; dCNRS, UMR 8619, Orsay, France Corresponding authors. B. de Foresta: beatrice.de-foresta@cea.fr J. Gallay: jacques.gallay@u-psud.fr

similaire:

	Guerrier de la Lumière Online – # 221 «On s’habitue parfois à ce que l’on voit dans les films, et l’on finit par oublier la vraie histoire», dit un ami, tandis que nous...		Museum communication and social media : the connected museum
	Déf : maladie infectieuse, contagieuse, avec évolution chronique....		Bulletin de liaison trimestriel «Dix Stratégies de Manipulation» à travers les média. Nous la reproduisons ici. Elle détaille l'éventail, depuis la stratégie de...
	Veille media «A quoi bon arriver quand l'action est terminée» a débarqué à Bakou 11 jours après le début des événements. Le Collectif van vous...		Supplementary Table 1
	Conversations à propos d’une émission de télévision Hey Christine, j’ai pensé à toi hier. J’ai regardé un reportage sur les chats hier, toi qui aimes tellement les animaux !		Table des matières
	Table des matières		Table des matieres