Visualization of F-actin filaments by a fluorescently labeled nucleotide analogue.
The tightly-bound nucleotide of F-actin was replaced with 1,N6-etheno-adenosine ATP (ADP). An epi-fluorescence optical microscope was modified to visualize efficiently the fluorescent analogue with an excitation-maximum wavelength of 310 nm. This microscope permitted us to visualize single F-actin filaments in solution using the fluorescence of the strongly bound 1,N6-ethenoadenosine nucleotide. Exchange of the tightly-bound nucleotide of F-actin with a free nucleotide in solution at a high temperature was quantitatively estimated by this method, and the results showed good agreement with results from phosphate release measurements.
Nakayama
H
H
Kansai Advanced Research Center, Communications Research Laboratory, Kobe, Japan. nak@crl.go.jp
Yamaga
T
T
D000199:Actins / Q000648:ultrastructure*; D000241:Adenosine / Q000031:analogs & derivatives; D005456:Fluorescent Dyes; D008856:Microscopy, Fluorescence; D009711:Nucleotides
D000199:Actins; D005456:Fluorescent Dyes; D009711:Nucleotides; C007640:1,N(6)-ethenoadenosine; D000241:Adenosine