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Reconstitution of Functionalized Transmembrane Domains of Receptor Proteins into Biomimetic Membranes
journal contribution
posted on 2015-08-25, 00:00 authored by Daniel R. Scott, Vitalii Silin, Hirsh NandaFor integral membrane proteins, an
assessment of their structures
and interactions within a biomimetic lipid bilayer environment is
critical for evaluating their cellular function. Hydrophobic sequences
prevalent within transmembrane domains, however, make these proteins
susceptible to aggregation and, thus, create difficulties in examining
their structural and functional properties via canonical techniques.
Working exclusively with single-pass transmembrane (TM) segments of
bitopic membrane proteins, in the form of soluble peptides, bypasses
many of the pitfalls of full-length protein preparations while allowing
for the opportunity to examine the properties of TM domains within
biomimetic membrane environments. In this study, peptides mimicking
the TM domains of the epidermal growth factor receptor (EGFR) and
CD4 co-receptor, both cell-signaling surface receptors, have been
reconstituted into 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine
(POPC) lipid bilayers. The formation of their native α-helical
structures within vesicle membranes was observed from circular dichroism,
and full partition of the peptides into the membrane was demonstrated
by tryptophan fluorescence and neutron reflectivity (NR). Using an
engineered planar lipid bilayer system ideal for surface characterization
methods, such as surface plasmon resonance (SPR) and NR, the TM peptides,
functionalized with a N-terminal biotin tag, proved capable of “activating”
a membrane surface, as evidenced by the capture of streptavidin. On
the basis of these initial assessments, we anticipate these membrane-bound
peptides will provide a versatile platform for understanding the intricate
roles of receptor TM domains in cell signaling.
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POPCTM domainsbiomimetic lipid bilayer environmentlipid bilayer systemSPRpeptideepidermal growth factor receptorsurface characterization methodsbitopic membrane proteinsbiomimetic membrane environmentsNRsurface plasmon resonanceEGFRreceptor TM domainsFunctionalized Transmembrane DomainsBiomimetic MembranesFor integral membrane proteins
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