TY - JOUR
T1 - Tuning membrane protein mobility by confinement into nanodomains
AU - Karner, Andreas
AU - Nimmervoll, Benedikt
AU - Plochberger, Birgit
AU - Klotzsch, Enrico
AU - Horner, Andreas
AU - Knyazev, Denis
AU - Kuttner, Roland
AU - Winkler, Klemens
AU - Winter, Lukas
AU - Siligan, Christine
AU - Ollinger, Nicole
AU - Pohl, Peter
AU - Preiner, Johannes
PY - 2017/3/7
Y1 - 2017/3/7
N2 - High-speed atomic force microscopy (HS-AFM) can be used to visualize function-related conformational changes of single
soluble proteins. Similar studies of single membrane proteins are, however, hampered by a lack of suitable flat, noninteracting
membrane supports and by high protein mobility. Here we show that streptavidin crystals grown on micasupported
lipid bilayers can be used as porous supports for membranes containing biotinylated lipids. Using SecYEG
(protein translocation channel) and GlpF (aquaglyceroporin), we demonstrate that the platform can be used to tune the
lateral mobility of transmembrane proteins to any value within the dynamic range accessible to HS-AFM imaging through
glutaraldehyde-cross-linking of the streptavidin. This allows HS-AFM to study the conformation or docking of spatially
confined proteins, which we illustrate by imaging GlpF at sub-molecular resolution and by observing the motor protein
SecA binding to SecYEG.
AB - High-speed atomic force microscopy (HS-AFM) can be used to visualize function-related conformational changes of single
soluble proteins. Similar studies of single membrane proteins are, however, hampered by a lack of suitable flat, noninteracting
membrane supports and by high protein mobility. Here we show that streptavidin crystals grown on micasupported
lipid bilayers can be used as porous supports for membranes containing biotinylated lipids. Using SecYEG
(protein translocation channel) and GlpF (aquaglyceroporin), we demonstrate that the platform can be used to tune the
lateral mobility of transmembrane proteins to any value within the dynamic range accessible to HS-AFM imaging through
glutaraldehyde-cross-linking of the streptavidin. This allows HS-AFM to study the conformation or docking of spatially
confined proteins, which we illustrate by imaging GlpF at sub-molecular resolution and by observing the motor protein
SecA binding to SecYEG.
KW - Escherichia coli/chemistry
KW - Escherichia coli Proteins/chemistry
KW - Lipid Bilayers/chemistry
KW - Membrane Proteins/chemistry
KW - Microscopy, Atomic Force/methods
KW - Protein Domains
KW - Protein Transport
UR - http://www.scopus.com/inward/record.url?scp=85008622419&partnerID=8YFLogxK
U2 - 10.1038/nnano.2016.236
DO - 10.1038/nnano.2016.236
M3 - Article
C2 - 27842062
SN - 1748-3387
VL - 12
SP - 260
EP - 266
JO - Nature Nanotechnology
JF - Nature Nanotechnology
IS - 3
ER -