Direct determination of oligomeric organization of integral membrane proteins and lipids from intact customizable bilayer

Aniruddha Panda; Fabian Giska; Anna L. Duncan; Alexander J. Welch; Caroline Brown; Rachel McAllister; Parameswaran Hariharan; Jean N.D. Goder; Jeff Coleman; Sathish Ramakrishnan; Frédéric Pincet; Lan Guan; Shyam Krishnakumar; James E. Rothman; Kallol Gupta

doi:10.1038/s41592-023-01864-5

Direct determination of oligomeric organization of integral membrane proteins and lipids from intact customizable bilayer

Aniruddha Panda, Fabian Giska, Anna L. Duncan, Alexander J. Welch, Caroline Brown, Rachel McAllister, Parameswaran Hariharan, Jean N.D. Goder, Jeff Coleman, Sathish Ramakrishnan, Frédéric Pincet, Lan Guan, Shyam Krishnakumar, James E. Rothman, Kallol Gupta^*

^*Corresponding author for this work

Department of Chemistry

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

18 Citations (Scopus)

Abstract

Hierarchical organization of integral membrane proteins (IMP) and lipids at the membrane is essential for regulating myriad downstream signaling. A quantitative understanding of these processes requires both detections of oligomeric organization of IMPs and lipids directly from intact membranes and determination of key membrane components and properties that regulate them. Addressing this, we have developed a platform that enables native mass spectrometry (nMS) analysis of IMP–lipid complexes directly from intact and customizable lipid membranes. Both the lipid composition and membrane properties (such as curvature, tension, and fluidity) of these bilayers can be precisely customized to a target membrane. Subsequent direct nMS analysis of these intact proteolipid vesicles can yield the oligomeric states of the embedded IMPs, identify bound lipids, and determine the membrane properties that can regulate the observed IMP–lipid organization. Applying this method, we show how lipid binding regulates neurotransmitter release and how membrane composition regulates the functional oligomeric state of a transporter.

Original language	English
Journal	Nature Methods
Volume	20
Issue	6
Pages (from-to)	891-897
Number of pages	7
ISSN	1548-7091
DOIs	https://doi.org/10.1038/s41592-023-01864-5
Publication status	Published - Jun 2023

Keywords

Biological Transport
Lipid Bilayers/chemistry
Lipids/chemistry
Mass Spectrometry/methods
Membrane Proteins/chemistry

Access to Document

10.1038/s41592-023-01864-5

https://pmc.ncbi.nlm.nih.gov/articles/PMC10932606/pdf/nihms-1972575.pdf

OpenUrl availability

Full text

Cite this

Panda, A., Giska, F., Duncan, A. L., Welch, A. J., Brown, C., McAllister, R., Hariharan, P., Goder, J. N. D., Coleman, J., Ramakrishnan, S., Pincet, F., Guan, L., Krishnakumar, S., Rothman, J. E., & Gupta, K. (2023). Direct determination of oligomeric organization of integral membrane proteins and lipids from intact customizable bilayer. Nature Methods, 20(6), 891-897. https://doi.org/10.1038/s41592-023-01864-5

@article{91b44790f7a246f0b98582f6fae03233,

title = "Direct determination of oligomeric organization of integral membrane proteins and lipids from intact customizable bilayer",

abstract = "Hierarchical organization of integral membrane proteins (IMP) and lipids at the membrane is essential for regulating myriad downstream signaling. A quantitative understanding of these processes requires both detections of oligomeric organization of IMPs and lipids directly from intact membranes and determination of key membrane components and properties that regulate them. Addressing this, we have developed a platform that enables native mass spectrometry (nMS) analysis of IMP–lipid complexes directly from intact and customizable lipid membranes. Both the lipid composition and membrane properties (such as curvature, tension, and fluidity) of these bilayers can be precisely customized to a target membrane. Subsequent direct nMS analysis of these intact proteolipid vesicles can yield the oligomeric states of the embedded IMPs, identify bound lipids, and determine the membrane properties that can regulate the observed IMP–lipid organization. Applying this method, we show how lipid binding regulates neurotransmitter release and how membrane composition regulates the functional oligomeric state of a transporter.",

keywords = "Biological Transport, Lipid Bilayers/chemistry, Lipids/chemistry, Mass Spectrometry/methods, Membrane Proteins/chemistry",

author = "Aniruddha Panda and Fabian Giska and Duncan, {Anna L.} and Welch, {Alexander J.} and Caroline Brown and Rachel McAllister and Parameswaran Hariharan and Goder, {Jean N.D.} and Jeff Coleman and Sathish Ramakrishnan and Fr{\'e}d{\'e}ric Pincet and Lan Guan and Shyam Krishnakumar and Rothman, {James E.} and Kallol Gupta",

year = "2023",

month = jun,

doi = "10.1038/s41592-023-01864-5",

language = "English",

volume = "20",

pages = "891--897",

journal = "Nature Methods",

issn = "1548-7091",

publisher = "Nature Research",

number = "6",

}

Panda, A, Giska, F, Duncan, AL, Welch, AJ, Brown, C, McAllister, R, Hariharan, P, Goder, JND, Coleman, J, Ramakrishnan, S, Pincet, F, Guan, L, Krishnakumar, S, Rothman, JE & Gupta, K 2023, 'Direct determination of oligomeric organization of integral membrane proteins and lipids from intact customizable bilayer', Nature Methods, vol. 20, no. 6, pp. 891-897. https://doi.org/10.1038/s41592-023-01864-5

Direct determination of oligomeric organization of integral membrane proteins and lipids from intact customizable bilayer. / Panda, Aniruddha; Giska, Fabian; Duncan, Anna L. et al.
In: Nature Methods, Vol. 20, No. 6, 06.2023, p. 891-897.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

TY - JOUR

T1 - Direct determination of oligomeric organization of integral membrane proteins and lipids from intact customizable bilayer

AU - Panda, Aniruddha

AU - Giska, Fabian

AU - Duncan, Anna L.

AU - Welch, Alexander J.

AU - Brown, Caroline

AU - McAllister, Rachel

AU - Hariharan, Parameswaran

AU - Goder, Jean N.D.

AU - Coleman, Jeff

AU - Ramakrishnan, Sathish

AU - Pincet, Frédéric

AU - Guan, Lan

AU - Krishnakumar, Shyam

AU - Rothman, James E.

AU - Gupta, Kallol

PY - 2023/6

Y1 - 2023/6

N2 - Hierarchical organization of integral membrane proteins (IMP) and lipids at the membrane is essential for regulating myriad downstream signaling. A quantitative understanding of these processes requires both detections of oligomeric organization of IMPs and lipids directly from intact membranes and determination of key membrane components and properties that regulate them. Addressing this, we have developed a platform that enables native mass spectrometry (nMS) analysis of IMP–lipid complexes directly from intact and customizable lipid membranes. Both the lipid composition and membrane properties (such as curvature, tension, and fluidity) of these bilayers can be precisely customized to a target membrane. Subsequent direct nMS analysis of these intact proteolipid vesicles can yield the oligomeric states of the embedded IMPs, identify bound lipids, and determine the membrane properties that can regulate the observed IMP–lipid organization. Applying this method, we show how lipid binding regulates neurotransmitter release and how membrane composition regulates the functional oligomeric state of a transporter.

AB - Hierarchical organization of integral membrane proteins (IMP) and lipids at the membrane is essential for regulating myriad downstream signaling. A quantitative understanding of these processes requires both detections of oligomeric organization of IMPs and lipids directly from intact membranes and determination of key membrane components and properties that regulate them. Addressing this, we have developed a platform that enables native mass spectrometry (nMS) analysis of IMP–lipid complexes directly from intact and customizable lipid membranes. Both the lipid composition and membrane properties (such as curvature, tension, and fluidity) of these bilayers can be precisely customized to a target membrane. Subsequent direct nMS analysis of these intact proteolipid vesicles can yield the oligomeric states of the embedded IMPs, identify bound lipids, and determine the membrane properties that can regulate the observed IMP–lipid organization. Applying this method, we show how lipid binding regulates neurotransmitter release and how membrane composition regulates the functional oligomeric state of a transporter.

KW - Biological Transport

KW - Lipid Bilayers/chemistry

KW - Lipids/chemistry

KW - Mass Spectrometry/methods

KW - Membrane Proteins/chemistry

UR - http://www.scopus.com/inward/record.url?scp=85153769661&partnerID=8YFLogxK

U2 - 10.1038/s41592-023-01864-5

DO - 10.1038/s41592-023-01864-5

M3 - Journal article

C2 - 37106230

AN - SCOPUS:85153769661

SN - 1548-7091

VL - 20

SP - 891

EP - 897

JO - Nature Methods

JF - Nature Methods

IS - 6

ER -

Direct determination of oligomeric organization of integral membrane proteins and lipids from intact customizable bilayer

Abstract

Keywords

Access to Document

OpenUrl availability

Other files and links

Fingerprint

Cite this