Membrane matters: The impact of a nanodisc-bilayer or a detergent microenvironment on the properties of two eubacterial rhodopsins
| Autor: | Laura Opdam, Sean Frehan, Srividya Ganapathy, Que Chen, Huub J. M. de Groot, Klaas J. Hellingwerf, John T. M. Kennis, Willem J. de Grip, Yusaku Hontani |
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| Přispěvatelé: | Biophysics Photosynthesis/Energy, LaserLaB - Energy |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Rhodopsin Detergent micelles Photoresponse Phosphorylcholine Thioglucosides Detergents Lipid Bilayers Biophysics Trimer Random hexamer 010402 general chemistry Cyanobacteria 01 natural sciences Biochemistry 03 medical and health sciences All institutes and research themes of the Radboud University Medical Center Bacterial Proteins Membrane proteins Maltose Nanodisc Microbial rhodopsins Proteorhodopsin biology Chemistry Protein Stability Bilayer Cell Biology Thermal stability 0104 chemical sciences 030104 developmental biology Membrane Membrane protein biology.protein Nanoparticles SDG 6 - Clean Water and Sanitation Nanomedicine Radboud Institute for Molecular Life Sciences [Radboudumc 19] |
| Zdroj: | Ganapathy, S, Opdam, L, Hontani, Y, Frehan, S, Chen, Q, Hellingwerf, K J, de Groot, H J M, Kennis, J T M & de Grip, W J 2020, ' Membrane matters : The impact of a nanodisc-bilayer or a detergent microenvironment on the properties of two eubacterial rhodopsins ', Biochimica et Biophysica Acta-Biomembranes, vol. 1862, no. 2, 183113, pp. 1-13 . https://doi.org/10.1016/j.bbamem.2019.183113 Biochimica et Biophysica Acta. Biomembranes, 1862 BBA-Biomembranes, 1862(2), 183113 BBA-Biomembranes Biochimica et Biophysica Acta-Biomembranes, 1862(2):183113, 1-13. Elsevier Biochimica et Biophysica Acta. Biomembranes, 1862, 2 Biochimica et Biophysica Acta-Biomembranes, 1862(2) |
| ISSN: | 1879-2642 0005-2736 |
| DOI: | 10.1016/j.bbamem.2019.183113 |
| Popis: | Multi-spanning membrane proteins usually require solubilization to allow proper purification and characterization, which generally impairs their structural and functional integrity. We have tested the efficacy of several commonly used detergents and membrane-mimicking nanodiscs with respect to solubilization, spectral properties, thermal stability and oligomeric profile of two membrane proteins from the eubacterial rhodopsin family, green proteorhodopsin (PR) and Gloeobacter violaceus rhodopsin (GR). Good solubilization was observed for the detergents TritonX-100 and dodecylphosphocholine (DPC), but DPC in particular strongly affected the thermal stability of PR and especially GR. The least deleterious effects were obtained with n-dodecyl-β-D-maltopyranoside (DDM) and octyl glucose neopentyl glycol (OGNG), which adequately stabilized the native oligomeric and monomeric state of PR and GR, respectively. The transition from the oligomeric to the monomeric state is accompanied by a small red-shift. Both GR and PR were rather unstable in SMA-nanodiscs, but the highest thermal stability was realized by the MSP-nanodisc environment. The size of the MSP-nanodisc was too small to fit the PR hexamer, but large enough to contain the PR monomer and GR trimer. This permitted the comparison of the photocycle of trimeric GR in a membrane-mimicking (MSP-nanodisc) and a detergent (DDM) environment. The ultrarapid early phase of the photocycle (femto- to picosecond lifetimes) showed very similar kinetics in either environment, but the slower part, initiated with proton transfer and generation of the M intermediate, proceeded faster in the nanodisc environment. The implications of our results for the biophysical characterization of PR and GR are discussed. |
| Databáze: | OpenAIRE |
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