scholarly journals Assembly of Model Membrane Nanodiscs for Native Mass Spectrometry

2021 ◽  
Author(s):  
Marius Kostelic ◽  
Ciara K. Zak ◽  
Hiruni Jayasekera ◽  
Michael Marty
Keyword(s):  
Mass Spectrometry ◽  
Membrane Protein ◽  
Lipid Bilayers ◽  
Biological Membranes ◽  
Native Mass Spectrometry ◽  
Model Membrane ◽  
Peptide Interactions ◽  
Lipid Nanodiscs ◽  
Natural Lipid ◽  
Selection Of

Native mass spectrometry (MS) with nanodiscs is a promising technique for characterizing membrane protein and peptide interactions in lipid bilayers. However, prior studies have used nanodiscs made of only one or two lipids, which lack the complexity of a natural lipid bilayer. To better model specific biological membranes, we developed model mammalian, bacterial, and mitochondrial nanodiscs with up to four different phospholipids. Careful selection of lipids with similar masses that balance the fluidity and curvature enabled these complex nanodiscs to be assembled and resolved with native MS. We then applied this approach to characterize the specificity and incorporation of LL-37, a human antimicrobial peptide, in single lipid nanodiscs versus model bacterial nanodiscs. Overall, development of these model membrane nanodiscs reveals new insights into the assembly of complex nanodiscs and provides a useful toolkit for studying membrane protein, peptide, and lipid interactions in model biological membranes.

scholarly journals Assembly of Model Membrane Nanodiscs for Native Mass Spectrometry

2021 ◽  
Author(s):  
Marius Kostelic ◽  
Ciara K. Zak ◽  
Hiruni Jayasekera ◽  
Michael Marty
Keyword(s):  
Mass Spectrometry ◽  
Membrane Protein ◽  
Lipid Bilayers ◽  
Biological Membranes ◽  
Native Mass Spectrometry ◽  
Model Membrane ◽  
Peptide Interactions ◽  
Lipid Nanodiscs ◽  
Natural Lipid ◽  
Selection Of

Native mass spectrometry (MS) with nanodiscs is a promising technique for characterizing membrane protein and peptide interactions in lipid bilayers. However, prior studies have used nanodiscs made of only one or two lipids, which lack the complexity of a natural lipid bilayer. To better model specific biological membranes, we developed model mammalian, bacterial, and mitochondrial nanodiscs with up to four different phospholipids. Careful selection of lipids with similar masses that balance the fluidity and curvature enabled these complex nanodiscs to be assembled and resolved with native MS. We then applied this approach to characterize the specificity and incorporation of LL-37, a human antimicrobial peptide, in single lipid nanodiscs versus model bacterial nanodiscs. Overall, development of these model membrane nanodiscs reveals new insights into the assembly of complex nanodiscs and provides a useful toolkit for studying membrane protein, peptide, and lipid interactions in model biological membranes.

Membrane Protein–Lipid Interactions Probed Using Mass Spectrometry

2019 ◽  
Vol 88 (1) ◽  
pp. 85-111 ◽  
Author(s):  
Jani Reddy Bolla ◽  
Mark T. Agasid ◽  
Shahid Mehmood ◽  
Carol V. Robinson
Keyword(s):  
Mass Spectrometry ◽  
Membrane Protein ◽  
Lipid Bilayers ◽  
Protein Interactions ◽  
Native Mass Spectrometry ◽  
Lipid Interactions ◽  
X Ray Crystallography ◽  
Native Ms ◽  
Molecular Aspects ◽  
Lipid Protein Interactions

Membrane proteins that exist in lipid bilayers are not isolated molecular entities. The lipid molecules that surround them play crucial roles in maintaining their full structural and functional integrity. Research directed at investigating these critical lipid–protein interactions is developing rapidly. Advancements in both instrumentation and software, as well as in key biophysical and biochemical techniques, are accelerating the field. In this review, we provide a brief outline of structural techniques used to probe protein–lipid interactions and focus on the molecular aspects of these interactions obtained from native mass spectrometry (native MS). We highlight examples in which lipids have been shown to modulate membrane protein structure and show how native MS has emerged as a complementary technique to X-ray crystallography and cryo–electron microscopy. We conclude with a short perspective on future developments that aim to better understand protein–lipid interactions in the native environment.

scholarly journals Native Mass Spectrometry of Antimicrobial Peptides in Lipid Nanodiscs Elucidates Complex Assembly

2019 ◽  
Vol 91 (14) ◽  
pp. 9284-9291 ◽  
Author(s):  
Lawrence R. Walker ◽  
Elaine M. Marzluff ◽  
Julia A. Townsend ◽  
William C. Resager ◽  
Michael T. Marty
Keyword(s):  
Mass Spectrometry ◽  
Antimicrobial Peptides ◽  
Native Mass Spectrometry ◽  
Complex Assembly ◽  
Lipid Nanodiscs

scholarly journals Scratching the surface: native mass spectrometry of peripheral membrane protein complexes

2020 ◽  
Vol 48 (2) ◽  
pp. 547-558 ◽  
Author(s):  
Cagla Sahin ◽  
Deseree J. Reid ◽  
Michael T. Marty ◽  
Michael Landreh
Keyword(s):  
Mass Spectrometry ◽  
Membrane Proteins ◽  
Membrane Protein ◽  
Lipid Vesicles ◽  
Native Mass Spectrometry ◽  
Integral Membrane Proteins ◽  
Structural Basis ◽  
Lipid Interactions ◽  
Native Ms ◽  
Peripheral Membrane Protein

A growing number of integral membrane proteins have been shown to tune their activity by selectively interacting with specific lipids. The ability to regulate biological functions via lipid interactions extends to the diverse group of proteins that associate only peripherally with the lipid bilayer. However, the structural basis of these interactions remains challenging to study due to their transient and promiscuous nature. Recently, native mass spectrometry has come into focus as a new tool to investigate lipid interactions in membrane proteins. Here, we outline how the native MS strategies developed for integral membrane proteins can be applied to generate insights into the structure and function of peripheral membrane proteins. Specifically, native MS studies of proteins in complex with detergent-solubilized lipids, bound to lipid nanodiscs, and released from native-like lipid vesicles all shed new light on the role of lipid interactions. The unique ability of native MS to capture and interrogate protein–protein, protein–ligand, and protein–lipid interactions opens exciting new avenues for the study of peripheral membrane protein biology.

Determining Membrane Protein–Lipid Binding Thermodynamics Using Native Mass Spectrometry

2016 ◽  
Vol 138 (13) ◽  
pp. 4346-4349 ◽  
Author(s):  
Xiao Cong ◽  
Yang Liu ◽  
Wen Liu ◽  
Xiaowen Liang ◽  
David H. Russell ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Membrane Protein ◽  
Native Mass Spectrometry ◽  
Lipid Binding ◽  
Binding Thermodynamics

scholarly journals Lipid binding attenuates channel closure of the outer membrane protein OmpF

2018 ◽  
Vol 115 (26) ◽  
pp. 6691-6696 ◽  
Author(s):  
Idlir Liko ◽  
Matteo T. Degiacomi ◽  
Sejeong Lee ◽  
Thomas D. Newport ◽  
Joseph Gault ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Membrane Protein ◽  
Outer Membrane ◽  
Lipid Bilayers ◽  
Protein Structures ◽  
Anion Channel ◽  
Lipid Binding ◽  
Coarse Grained ◽  
Strong Interactions ◽  
Voltage Dependent Anion Channel

Strong interactions between lipids and proteins occur primarily through association of charged headgroups and amino acid side chains, rendering the protonation status of both partners important. Here we use native mass spectrometry to explore lipid binding as a function of charge of the outer membrane porin F (OmpF). We find that binding of anionic phosphatidylglycerol (POPG) or zwitterionic phosphatidylcholine (POPC) to OmpF is sensitive to electrospray polarity while the effects of charge are less pronounced for other proteins in outer or mitochondrial membranes: the ferripyoverdine receptor (FpvA) or the voltage-dependent anion channel (VDAC). Only marginal charge-induced differences were observed for inner membrane proteins: the ammonia channel (AmtB) or the mechanosensitive channel. To understand these different sensitivities, we performed an extensive bioinformatics analysis of membrane protein structures and found that OmpF, and to a lesser extent FpvA and VDAC, have atypically high local densities of basic and acidic residues in their lipid headgroup-binding regions. Coarse-grained molecular dynamics simulations, in mixed lipid bilayers, further implicate changes in charge by demonstrating preferential binding of anionic POPG over zwitterionic POPC to protonated OmpF, an effect not observed to the same extent for AmtB. Moreover, electrophysiology and mass-spectrometry–based ligand-binding experiments, at low pH, show that POPG can maintain OmpF channels in open conformations for extended time periods. Since the outer membrane is composed almost entirely of anionic lipopolysaccharide, with similar headgroup properties to POPG, such anionic lipid binding could prevent closure of OmpF channels, thereby increasing access of antibiotics that use porin-mediated pathways.

Assembly of Model Membrane Nanodiscs for Native Mass Spectrometry

2021 ◽  
Author(s):  
Marius M. Kostelic ◽  
Ciara K. Zak ◽  
Hiruni S. Jayasekera ◽  
Michael T. Marty
Keyword(s):  
Mass Spectrometry ◽  
Native Mass Spectrometry ◽  
Model Membrane

Exploration of interactions between membrane proteins embedded in supported lipid bilayers and their antibodies by reflectometric interference spectroscopy-based sensing

The Analyst ◽  
2014 ◽  
Vol 139 (22) ◽  
pp. 6016-6021 ◽  
Author(s):  
Yoshikazu Kurihara ◽  
Tsuneo Sawazumi ◽  
Toshifumi Takeuchi
Keyword(s):  
Multidrug Resistance ◽  
Membrane Proteins ◽  
Membrane Protein ◽  
Lipid Bilayers ◽  
Model Membrane ◽  
Supported Lipid Bilayers ◽  
Reflectometric Interference Spectroscopy

A microfluidic reflectometric interference spectroscopy (RIfS)-based sensor was fabricated to investigate the activity of multidrug resistance-associated protein 1 (MRP1), applied as a model membrane protein.

Sign in / Sign up

Export Citation Format

Share Document