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Proc Natl Acad Sci U S A
2002 Oct 01;9920:12759-64. doi: 10.1073/pnas.192261499.
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Structural rearrangements in single ion channels detected optically in living cells.
Sonnleitner A, Mannuzzu LM, Terakawa S, Isacoff EY.
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Total internal reflection fluorescence microscopy was used to detect single fluorescently labeled voltage-gated Shaker K(+) channels in the plasma membrane of living cells. Tetramethylrhodamine (TMR) attached to specific amino acid positions in the voltage-sensing S4 segment changed fluorescence intensity in response to the voltage-driven protein motions of the channel. The voltage dependence of the fluorescence of single TMRs was similar to that seen in macroscopic epi-illumination microscopy, but the exclusion of nonchannel fluorescence revealed that the dimming of TMR upon voltage sensor rearrangement was much larger than previously thought, and is due to an extreme, approximately 20-fold suppression of the elementary fluorescence. The total internal reflection voltage-clamp method reveals protein motions that do not directly open or close the ion channel and which have therefore not been detected before at the single-channel level. The method should be applicable to a wide assortment of membrane-associated proteins and should make it possible to relate the structural rearrangements of single proteins to simultaneously measured physiological cell-signaling events.
Axelrod,
Total internal reflection fluorescence microscopy.
1989, Pubmed
Axelrod,
Total internal reflection fluorescence microscopy.
1989,
Pubmed Baker,
Three transmembrane conformations and sequence-dependent displacement of the S4 domain in shaker K+ channel gating.
1998,
Pubmed Bluemink,
Freeze-fracture electron microscopy of membrane changes in progesterone-induced maturing oocytes and eggs of Xenopus laevis.
1983,
Pubmed
,
Xenbase Cha,
Atomic scale movement of the voltage-sensing region in a potassium channel measured via spectroscopy.
1999,
Pubmed Cha,
Structural implications of fluorescence quenching in the Shaker K+ channel.
1998,
Pubmed Cha,
Characterizing voltage-dependent conformational changes in the Shaker K+ channel with fluorescence.
1997,
Pubmed
,
Xenbase Funatsu,
Imaging of single fluorescent molecules and individual ATP turnovers by single myosin molecules in aqueous solution.
1995,
Pubmed Gandhi,
Reconstructing voltage sensor-pore interaction from a fluorescence scan of a voltage-gated K+ channel.
2000,
Pubmed Glauner,
Spectroscopic mapping of voltage sensor movement in the Shaker potassium channel.
1999,
Pubmed
,
Xenbase Hoshi,
Biophysical and molecular mechanisms of Shaker potassium channel inactivation.
1990,
Pubmed
,
Xenbase Kamb,
Molecular characterization of Shaker, a Drosophila gene that encodes a potassium channel.
1987,
Pubmed Kobilka,
Examination of ligand-induced conformational changes in the beta 2-adrenergic receptor by fluorescence spectroscopy.
1998,
Pubmed Larsson,
Transmembrane movement of the shaker K+ channel S4.
1996,
Pubmed
,
Xenbase Li,
An intermediate state of the gamma-aminobutyric acid transporter GAT1 revealed by simultaneous voltage clamp and fluorescence.
2000,
Pubmed
,
Xenbase Liu,
Site-directed fluorescence labeling of P-glycoprotein on cysteine residues in the nucleotide binding domains.
1996,
Pubmed Loots,
Protein rearrangements underlying slow inactivation of the Shaker K+ channel.
1998,
Pubmed Mannuzzu,
Independence and cooperativity in rearrangements of a potassium channel voltage sensor revealed by single subunit fluorescence.
2000,
Pubmed
,
Xenbase Mannuzzu,
Direct physical measure of conformational rearrangement underlying potassium channel gating.
1996,
Pubmed
,
Xenbase Mehta,
Single-molecule biomechanics with optical methods.
1999,
Pubmed Perozo,
Gating currents from a nonconducting mutant reveal open-closed conformations in Shaker K+ channels.
1993,
Pubmed Starace,
Voltage-dependent proton transport by the voltage sensor of the Shaker K+ channel.
1997,
Pubmed Steyer,
Transport, docking and exocytosis of single secretory granules in live chromaffin cells.
1997,
Pubmed Xie,
Single-molecule enzymology.
1999,
Pubmed Yang,
Molecular basis of charge movement in voltage-gated sodium channels.
1996,
Pubmed Yusaf,
Measurement of the movement of the S4 segment during the activation of a voltage-gated potassium channel.
1996,
Pubmed
,
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