TY - JOUR
T1 - Investigating Daptomycin-Membrane Interactions Using Native MS and Fast Photochemical Oxidation of Peptides in Nanodiscs
AU - Reid, Deseree J.
AU - Dash, Tapasyatanu
AU - Wang, Zhihan
AU - Aspinwall, Craig A.
AU - Marty, Michael T.
N1 - Funding Information: The analysis for calcium concentration in daptomycin was performed by Mary Kay Amistadi in the Arizona Laboratory for Emerging Contaminants at the University of Arizona, Tucson, AZ. The pMSP1D1 plasmid was a gift from Stephen Sligar (Addgene plasmid #20061). This work was funded by the National Institutes of Health (R35 GM128624) and the National Science Foundation (CBET-2003297). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health or the National Science Foundation. The authors thank Alexander Makarov, Mike Senko, Tobias Woerner, and Kyle Fort at Thermo Fisher Scientific for support on the UHMR Q-Exactive HF instrument. Publisher Copyright: © 2023 American Chemical Society.
PY - 2023/3/21
Y1 - 2023/3/21
N2 - Daptomycin is a cyclic lipopeptide antibiotic that targets the lipid membrane of Gram-positive bacteria. Membrane fluidity and charge can affect daptomycin activity, but its mechanisms are poorly understood because it is challenging to study daptomycin interactions within lipid bilayers. Here, we combined native mass spectrometry (MS) and fast photochemical oxidation of peptides (FPOP) to study daptomycin-membrane interactions with different lipid bilayer nanodiscs. Native MS suggests that daptomycin incorporates randomly and does not prefer any specific oligomeric states when integrated into bilayers. FPOP reveals significant protection in most bilayer environments. Combining the native MS and FPOP results, we observed that stronger membrane interactions are formed with more rigid membranes, and pore formation may occur in more fluid membranes to expose daptomycin to FPOP oxidation. Electrophysiology measurements further supported the observation of polydisperse pore complexes from the MS data. Together, these results demonstrate the complementarity of native MS, FPOP, and membrane conductance experiments to shed light on how antibiotic peptides interact with and within lipid membranes.
AB - Daptomycin is a cyclic lipopeptide antibiotic that targets the lipid membrane of Gram-positive bacteria. Membrane fluidity and charge can affect daptomycin activity, but its mechanisms are poorly understood because it is challenging to study daptomycin interactions within lipid bilayers. Here, we combined native mass spectrometry (MS) and fast photochemical oxidation of peptides (FPOP) to study daptomycin-membrane interactions with different lipid bilayer nanodiscs. Native MS suggests that daptomycin incorporates randomly and does not prefer any specific oligomeric states when integrated into bilayers. FPOP reveals significant protection in most bilayer environments. Combining the native MS and FPOP results, we observed that stronger membrane interactions are formed with more rigid membranes, and pore formation may occur in more fluid membranes to expose daptomycin to FPOP oxidation. Electrophysiology measurements further supported the observation of polydisperse pore complexes from the MS data. Together, these results demonstrate the complementarity of native MS, FPOP, and membrane conductance experiments to shed light on how antibiotic peptides interact with and within lipid membranes.
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U2 - https://doi.org/10.1021/acs.analchem.2c05222
DO - https://doi.org/10.1021/acs.analchem.2c05222
M3 - Article
SN - 0003-2700
VL - 95
SP - 4984
EP - 4991
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 11
ER -