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IEEE Trans Nanobioscience
2005 Dec 01;44:269-76. doi: 10.1109/tnb.2005.859552.
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Molecular handles for the mechanical manipulation of single-membrane proteins in living cells.
Gorostiza P
,
Tombola F
,
Verdaguer A
,
Smith SB
,
Bustamante C
,
Isacoff EY
.
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We have developed a procedure to selectively biotinylate a specific membrane protein, enabling its attachment to external force probes and thus allowing its mechanical manipulation within its native environment. Using potassium channels as model membrane proteins in oocytes, we have found that Maleimide-PEG3400-biotin is the crosslinker with highest conjugation selectivity and accessibility to external probes. Neutravidin-coated beads provide for directed attachment while avoiding nonspecific interactions with the cell. The technology was successfully tested by mechanical manipulation of biotinylated extracellular residues of channels in oocytes using an atomic force microscope under conditions which preserve function of the channels. Binding forces of approximately 80 pN at 100 nN/s were measured.
Afrin,
Analysis of force curves obtained on the live cell membrane using chemically modified AFM probes.
2004, Pubmed
Afrin,
Analysis of force curves obtained on the live cell membrane using chemically modified AFM probes.
2004,
Pubmed
Baker,
Three transmembrane conformations and sequence-dependent displacement of the S4 domain in shaker K+ channel gating.
1998,
Pubmed
Bustamante,
Mechanical processes in biochemistry.
2004,
Pubmed
Chen,
Single-molecule force measurements.
2002,
Pubmed
Friedsam,
Polymer functionalized AFM tips for long-term measurements in single-molecule force spectroscopy.
2004,
Pubmed
Hiller,
Biotin binding to avidin. Oligosaccharide side chain not required for ligand association.
1987,
Pubmed
Larsson,
Transmembrane movement of the shaker K+ channel S4.
1996,
Pubmed
,
Xenbase
Maier,
DNA transport into Bacillus subtilis requires proton motive force to generate large molecular forces.
2004,
Pubmed
Mannuzzu,
Direct physical measure of conformational rearrangement underlying potassium channel gating.
1996,
Pubmed
,
Xenbase
Raab,
Antibody recognition imaging by force microscopy.
1999,
Pubmed
Ratto,
Force spectroscopy of the double-tethered concanavalin-A mannose bond.
2004,
Pubmed
Ryu,
Torque-generating units of the flagellar motor of Escherichia coli have a high duty ratio.
2000,
Pubmed
Smith,
The bacteriophage straight phi29 portal motor can package DNA against a large internal force.
2001,
Pubmed
Sonnleitner,
Structural rearrangements in single ion channels detected optically in living cells.
2002,
Pubmed
,
Xenbase
