Abstract
Studying membrane proteins at the molecular level represents a major challenge in biochemistry due to the complexity of the membrane in which they are embedded. As an important step towards a detailed understanding of their action and molecular functioning, current studies focus on membrane proteins reconstituted into artificial lipid environments. Such reconstituted systems allow for a more flexible choice of biochemical, biophysical, and microscopy techniques for characterizing the proteins. This review gives an overview of the methods currently available for reconstituting membrane proteins in a functional state into giant unilamellar vesicles, and discusses some key methods to verify successful reconstitution.
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Abbreviations
- AFM:
-
Atomic force microscopy
- AC-electric field:
-
Alternate current-electric field
- BR:
-
Bacteriorhodopsin
- DIC:
-
Differential interference contrast
- GPMVs:
-
Giant plasma membrane vesicles
- GUVs:
-
Giant unilamellar vesicles
- ITO:
-
Indium tin oxide
- LUVs:
-
Large unilamellar vesicles
- PVA:
-
Polyvinyl alcohol
- SUVs:
-
Small unilamellar vesicles
- T m :
-
Miscibility temperature
- VDAC:
-
Voltage-dependent anion channel
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Acknowledgments
Research in the authors' lab was supported by the Danish Council for Independent Research | Natural Sciences (FNU, Grant Number 1323-00297), the Research Centre ‘bioSYNergy’ funded by the UCPH Excellence Programme for Interdisciplinary Research, the Villum Foundation (Project Number VKR023115) and the Danish National Research Foundation through the PUMPKIN Centre of Excellence (DNRF85). The anonymous reviewer is thanked for critically reading the manuscript and suggesting substantial improvements.
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Jørgensen, I.L., Kemmer, G.C. & Pomorski, T.G. Membrane protein reconstitution into giant unilamellar vesicles: a review on current techniques. Eur Biophys J 46, 103–119 (2017). https://doi.org/10.1007/s00249-016-1155-9
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DOI: https://doi.org/10.1007/s00249-016-1155-9