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Quantification of Protein-Induced Membrane Remodeling Kinetics In Vitro with Lipid Multilayer Gratings.

Title: Quantification of Protein-Induced Membrane Remodeling Kinetics In Vitro with Lipid Multilayer Gratings.
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Name(s): Lowry, Troy W, author
Hariri, Hanaa, author
Prommapan, Plengchart, author
Kusi-Appiah, Aubrey, author
Vafai, Nicholas, author
Bienkiewicz, Ewa A, author
Van Winkle, David H, author
Stagg, Scott M, author
Lenhert, Steven, author
Type of Resource: text
Genre: Journal Article
Text
Date Issued: 2016-01-27
Physical Form: computer
online resource
Extent: 1 online resource
Language(s): English
Abstract/Description: The dynamic self-organization of lipids in biological systems is a highly regulated process that enables the compartmentalization of living systems at micro- and nanoscopic scales. Consequently, quantitative methods for assaying the kinetics of supramolecular remodeling such as vesicle formation from planar lipid bilayers or multilayers are needed to understand cellular self-organization. Here, a new nanotechnology-based method for quantitative measurements of lipid-protein interactions is presented and its suitability for quantifying the membrane binding, inflation, and budding activity of the membrane-remodeling protein Sar1 is demonstrated. Lipid multilayer gratings are printed onto surfaces using nanointaglio and exposed to Sar1, resulting in the inflation of lipid multilayers into unilamellar structures, which can be observed in a label-free manner by monitoring the diffracted light. Local variations in lipid multilayer volume on the surface is used to vary substrate availability in a microarray format. A quantitative model is developed that allows quantification of binding affinity (K D ) and kinetics (kon and koff ). Importantly, this assay is uniquely capable of quantifying membrane remodeling. Upon Sar1-induced inflation of single bilayers from surface supported multilayers, the semicylindrical grating lines are observed to remodel into semispherical buds when a critical radius of curvature is reached.
Identifier: FSU_pmch_26649649 (IID), 10.1002/smll.201502398 (DOI), PMC4843995 (PMCID), 26649649 (RID), 26649649 (EID)
Keywords: Biosensors, Kinetics, Lipid multilayer gratings, Lipids, Membranes, Microarrays
Grant Number: R01 GM086892, R01 GM107172
Publication Note: This NIH-funded author manuscript originally appeared in PubMed Central at https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4843995.
Subject(s): Kinetics
Light
Lipid Bilayers/metabolism
Membrane Proteins/metabolism
Microscopy, Fluorescence
Optical Phenomena
Persistent Link to This Record: http://purl.flvc.org/fsu/fd/FSU_pmch_26649649
Owner Institution: FSU
Is Part Of: Small (Weinheim an der Bergstrasse, Germany).
1613-6829
Issue: iss. 4, vol. 12

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Lowry, T. W., Hariri, H., Prommapan, P., Kusi-Appiah, A., Vafai, N., Bienkiewicz, E. A., … Lenhert, S. (2016). Quantification of Protein-Induced Membrane Remodeling Kinetics In Vitro with Lipid Multilayer Gratings. Small (Weinheim An Der Bergstrasse, Germany). Retrieved from http://purl.flvc.org/fsu/fd/FSU_pmch_26649649