Abstract
Membrane-embedded proteins (MPs) are central to a wide range of cellular processes. Despite their importance, structural studies of MPs are hindered by expression difficulties and the need for stabilization in a membrane-mimicking environment. High-resolution NMR methods can investigate structure and function of MPs due to methodological advances and new membrane-like assemblies for stabilization of MPs. In this perspective of the field, we introduce the challenges and opportunities of NMR studies of membrane proteins, briefly surveying membrane-mimicking systems and their application in structure determination. A case study then focuses on the C-terminal domain of the bacterial potassium channel KcsA, describing how improvements in membrane-mimicking conditions eventually enabled us to present a structural view of the pH-dependent behavior of this cytoplasmic channel domain. The results highlight prerequisites for a successful study of MPs and the potential for future investigations.
| Original language | English |
|---|---|
| Pages (from-to) | 1001-1013 |
| Number of pages | 13 |
| Journal | Israel Journal of Chemistry |
| Volume | 59 |
| Issue number | 11-12 |
| DOIs | |
| State | Published - 1 Nov 2019 |
Bibliographical note
Publisher Copyright:© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
Funding
We are indebted to many co-workers contributing to the findings described in this paper, including Dr. John M. Louis (NIDDK, NIH, Bethesda, MD, USA), Dr. Guy Kamnesky (Technion – Israel Institute of Technology, Haifa, Israel), and Dr. Hadassa Shaked and Orel Hirschhorn (BIU). We are grateful to Prof. Sharon Ruthstein and Zena Qasem for performing and analyzing the CW-EPR experiments. The guidance of Dr. Adriaan Bax (NIDDK, NIH, Bethesda, MD, USA) in the critical initial stages of this study is greatly appreciated. We also thank Drs. Hugo Gottlieb and Keren Keinan-Adamsky for spectrometer assistance and Israel Tabakman for technical assistance. The MSP1D1-encoding plasmid (for preparation of LPNs) was generously provided by Prof. Franz Hagn (Technische Universitat München). Financial support by a long-term EMBO fellowship and an NIH-Director fellowship (to JHC) and funding by the Israel Science Foundation (award 1088/16) and the Binational Israel-US Foundation (awards 2013185 and 2017243) is gratefully acknowledged. Establishment of the 700 MHz spectrometer system was supported by Fundácion Adar and a Converging Technologies award. JHC acknowledges support of the Christians for Israel Chair for Medical Research. We are indebted to many co‐workers contributing to the findings described in this paper, including Dr. John M. Louis (NIDDK, NIH, Bethesda, MD, USA), Dr. Guy Kamnesky (Technion – Israel Institute of Technology, Haifa, Israel), and Dr. Hadassa Shaked and Orel Hirschhorn (BIU). We are grateful to Prof. Sharon Ruthstein and Zena Qasem for performing and analyzing the CW‐EPR experiments. The guidance of Dr. Adriaan Bax (NIDDK, NIH, Bethesda, MD, USA) in the critical initial stages of this study is greatly appreciated. We also thank Drs. Hugo Gottlieb and Keren Keinan‐Adamsky for spectrometer assistance and Israel Tabakman for technical assistance. The MSP1D1‐encoding plasmid (for preparation of LPNs) was generously provided by Prof. Franz Hagn (Technische Universitat München). Financial support by a long‐term EMBO fellowship and an NIH‐Director fellowship (to JHC) and funding by the Israel Science Foundation (award 1088/16) and the Binational Israel‐US Foundation ( awards 2013185 and 2017243) is gratefully acknowledged. Establishment of the 700 MHz spectrometer system was supported by Fundácion Adar and a Converging Technologies award. JHC acknowledges support of the Christians for Israel Chair for Medical Research.
| Funders | Funder number |
|---|---|
| Binational Israel-US Foundation | |
| Binational Israel‐US Foundation | 2017243, 2013185 |
| NIH-Director | |
| Technion – Israel Institute of Technology, Haifa, Israel | |
| National Institutes of Health | |
| National Institute of Diabetes and Digestive and Kidney Diseases | |
| European Molecular Biology Organization | |
| Israel Science Foundation | 1088/16 |
Keywords
- Electron paramagnetic resonance
- KcsA channel
- Lipoprotein nanodiscs
- Membrane proteins
- Nuclear magnetic resonance