Biophysical Characterization of Membrane Proteins Embedded in Nanodiscs Using Fluorescence Correlation Spectroscopy
Abstract
Proteins embedded in biological membranes perform essential functions in all organisms, serving as receptors, transporters, channels, cell adhesion molecules, and other supporting cellular roles. These membrane proteins comprise ~30% of all human proteins and are the targets of ~60% of FDA-approved drugs, yet their extensive characterization using established biochemical and biophysical methods has continued to be elusive due to challenges associated with the purification of these insoluble proteins. In response, the development of nanodisc techniques, such as nanolipoprotein particles (NLPs) and styrene maleic acid polymers (SMALPs), allowed membrane proteins to be expressed and isolated in solution as part of lipid bilayer rafts with defined, consistent nanometer sizes and compositions, thus enabling solution-based measurements. Fluorescence correlation spectroscopy (FCS) is a relatively simple yet powerful optical microscopy-based technique that yields quantitative biophysical information, such as diffusion kinetics and concentrations, about individual or interacting species in solution. Here, we first summarize current nanodisc techniques and FCS fundamentals. We then provide a focused review of studies that employed FCS in combination with nanodisc technology to investigate a handful of membrane proteins, including bacteriorhodopsin, bacterial division protein ZipA, bacterial membrane insertases SecYEG and YidC, Yersinia pestis type III secretion protein YopB, yeast cell wall stressmore »
- Publication Date:
- Research Org.:
- Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
- Sponsoring Org.:
- USDOE National Nuclear Security Administration (NNSA); National Institutes of Health (NIH); USDOE Laboratory Directed Research and Development (LDRD) Program. Lawrence Livermore National Laboratory (LLNL)
- OSTI Identifier:
- 1860531
- Alternate Identifier(s):
- OSTI ID: 1872282
- Report Number(s):
- LLNL-JRNL-836103
Journal ID: ISSN 2077-0375; MBSEB6; PII: membranes12040392
- Grant/Contract Number:
- AC52-07NA27344; Laboratory Directed Research and Development; 1R01GM117342; U19AI144184
- Resource Type:
- Published Article
- Journal Name:
- Membranes
- Additional Journal Information:
- Journal Name: Membranes Journal Volume: 12 Journal Issue: 4; Journal ID: ISSN 2077-0375
- Publisher:
- MDPI AG
- Country of Publication:
- Switzerland
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; fluorescent correlation spectroscopy; membrane proteins; nanodiscs; cell-free expression
Citation Formats
Laurence, Matthew J., Carpenter, Timothy S., Laurence, Ted A., Coleman, Matthew A., Shelby, Megan, and Liu, Chao. Biophysical Characterization of Membrane Proteins Embedded in Nanodiscs Using Fluorescence Correlation Spectroscopy. Switzerland: N. p., 2022.
Web. doi:10.3390/membranes12040392.
Laurence, Matthew J., Carpenter, Timothy S., Laurence, Ted A., Coleman, Matthew A., Shelby, Megan, & Liu, Chao. Biophysical Characterization of Membrane Proteins Embedded in Nanodiscs Using Fluorescence Correlation Spectroscopy. Switzerland. https://doi.org/10.3390/membranes12040392
Laurence, Matthew J., Carpenter, Timothy S., Laurence, Ted A., Coleman, Matthew A., Shelby, Megan, and Liu, Chao. Thu .
"Biophysical Characterization of Membrane Proteins Embedded in Nanodiscs Using Fluorescence Correlation Spectroscopy". Switzerland. https://doi.org/10.3390/membranes12040392.
@article{osti_1860531,
title = {Biophysical Characterization of Membrane Proteins Embedded in Nanodiscs Using Fluorescence Correlation Spectroscopy},
author = {Laurence, Matthew J. and Carpenter, Timothy S. and Laurence, Ted A. and Coleman, Matthew A. and Shelby, Megan and Liu, Chao},
abstractNote = {Proteins embedded in biological membranes perform essential functions in all organisms, serving as receptors, transporters, channels, cell adhesion molecules, and other supporting cellular roles. These membrane proteins comprise ~30% of all human proteins and are the targets of ~60% of FDA-approved drugs, yet their extensive characterization using established biochemical and biophysical methods has continued to be elusive due to challenges associated with the purification of these insoluble proteins. In response, the development of nanodisc techniques, such as nanolipoprotein particles (NLPs) and styrene maleic acid polymers (SMALPs), allowed membrane proteins to be expressed and isolated in solution as part of lipid bilayer rafts with defined, consistent nanometer sizes and compositions, thus enabling solution-based measurements. Fluorescence correlation spectroscopy (FCS) is a relatively simple yet powerful optical microscopy-based technique that yields quantitative biophysical information, such as diffusion kinetics and concentrations, about individual or interacting species in solution. Here, we first summarize current nanodisc techniques and FCS fundamentals. We then provide a focused review of studies that employed FCS in combination with nanodisc technology to investigate a handful of membrane proteins, including bacteriorhodopsin, bacterial division protein ZipA, bacterial membrane insertases SecYEG and YidC, Yersinia pestis type III secretion protein YopB, yeast cell wall stress sensor Wsc1, epidermal growth factor receptor (EGFR), ABC transporters, and several G protein-coupled receptors (GPCRs).},
doi = {10.3390/membranes12040392},
journal = {Membranes},
number = 4,
volume = 12,
place = {Switzerland},
year = {Thu Mar 31 00:00:00 EDT 2022},
month = {Thu Mar 31 00:00:00 EDT 2022}
}
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