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2015 ; 564
(ä): 219-58
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Peptide-Membrane Interactions by Spin-Labeling EPR
#MMPMID26477253
Smirnova TI
; Smirnov AI
Methods Enzymol
2015[]; 564
(ä): 219-58
PMID26477253
show ga
Site-directed spin labeling (SDSL) in combination with electron paramagnetic
resonance (EPR) spectroscopy is a well-established method that has recently grown
in popularity as an experimental technique, with multiple applications in protein
and peptide science. The growth is driven by development of labeling strategies,
as well as by considerable technical advances in the field, that are paralleled
by an increased availability of EPR instrumentation. While the method requires an
introduction of a paramagnetic probe at a well-defined position in a peptide
sequence, it has been shown to be minimally destructive to the peptide structure
and energetics of the peptide-membrane interactions. In this chapter, we describe
basic approaches for using SDSL EPR spectroscopy to study interactions between
small peptides and biological membranes or membrane mimetic systems. We focus on
experimental approaches to quantify peptide-membrane binding, topology of bound
peptides, and characterize peptide aggregation. Sample preparation protocols
including spin-labeling methods and preparation of membrane mimetic systems are
also described.
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