The Motion of a Single Molecule, the Lambda-Receptor, in the Bacterial Outer Membrane
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The Motion of a Single Molecule, the Lambda-Receptor, in the Bacterial Outer Membrane. / Oddershede, Lene; Dreyer, Jakob Kisbye; Grego, Sonia; Brown, Stanley; Berg-Sørensen, Kirstine.
I: Biophysical Journal, Bind 83, Nr. 6, 2002, s. 3152-61.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - The Motion of a Single Molecule, the Lambda-Receptor, in the Bacterial Outer Membrane
AU - Oddershede, Lene
AU - Dreyer, Jakob Kisbye
AU - Grego, Sonia
AU - Brown, Stanley
AU - Berg-Sørensen, Kirstine
N1 - Keywords: Bacterial Outer Membrane Proteins; Biotin; Biotinylation; Computer Simulation; Elasticity; Escherichia coli; Lasers; Micromanipulation; Microspheres; Models, Biological; Models, Chemical; Motion; Nanotechnology; Optics and Photonics; Particle Size; Porins; Protein Conformation; Receptors, Virus; Recombinant Proteins; Streptavidin; Stress, Mechanical
PY - 2002
Y1 - 2002
N2 - Using optical tweezers and single particle tracking, we have revealed the motion of a single protein, the lambda-receptor, in the outer membrane of living Escherichia coli bacteria. We genetically modified the lambda-receptor placing a biotin on an extracellular site of the receptor in vivo. The efficiency of this in vivo biotinylation is very low, thus enabling the attachment of a streptavidin-coated bead binding specifically to a single biotinylated lambda-receptor. The bead was used as a handle for the optical tweezers and as a marker for the single particle tracking routine. We propose a model that allows extraction of the motion of the protein from measurements of the mobility of the bead-molecule complex; these results are equally applicable to analyze bead-protein complexes in other membrane systems. Within a domain of radius approximately 25 nm, the receptor diffuses with a diffusion constant of (1.5 +/- 1.0) x 10(-9) cm(2)/s and sits in a harmonic potential as if it were tethered by an elastic spring of spring constant of ~1.0 x 10(-2) pN/nm to the bacterial membrane. The purpose of the protein motion might be to facilitate transport of maltodextrins through the outer bacterial membrane.
AB - Using optical tweezers and single particle tracking, we have revealed the motion of a single protein, the lambda-receptor, in the outer membrane of living Escherichia coli bacteria. We genetically modified the lambda-receptor placing a biotin on an extracellular site of the receptor in vivo. The efficiency of this in vivo biotinylation is very low, thus enabling the attachment of a streptavidin-coated bead binding specifically to a single biotinylated lambda-receptor. The bead was used as a handle for the optical tweezers and as a marker for the single particle tracking routine. We propose a model that allows extraction of the motion of the protein from measurements of the mobility of the bead-molecule complex; these results are equally applicable to analyze bead-protein complexes in other membrane systems. Within a domain of radius approximately 25 nm, the receptor diffuses with a diffusion constant of (1.5 +/- 1.0) x 10(-9) cm(2)/s and sits in a harmonic potential as if it were tethered by an elastic spring of spring constant of ~1.0 x 10(-2) pN/nm to the bacterial membrane. The purpose of the protein motion might be to facilitate transport of maltodextrins through the outer bacterial membrane.
U2 - 10.1016/S0006-3495(02)75318-6
DO - 10.1016/S0006-3495(02)75318-6
M3 - Journal article
C2 - 12496085
VL - 83
SP - 3152
EP - 3161
JO - Biophysical Journal
JF - Biophysical Journal
SN - 0006-3495
IS - 6
ER -
ID: 128538