Reconstitution of the muscle thin filament from recombinant troponin components and the native thin filaments.
We have developed a technique by which muscle thin filaments are reconstituted from the recombinant troponin components and the native thin filaments. By this technique, the reconstituted troponin complex is exchanged into the native thin filaments in the presence of 20% glycerol and 0.3M KCl at pH 6.2. More than 90% of endogenous troponin complex was replaced with the recombinant troponin complex. Structural integrity and Ca(2+) sensitivity of the reconstituted thin filament prepared by this technique was confirmed by X-ray fiber diffraction measurements and the thin filament-activated myosin subfragment 1 ATPase measurements, respectively.
Matsumoto
Fumiko
F
Quantum Beam Science Directorate, Japan Atomic Energy Agency, Naka-gun, Ibaraki 319-1195, Japan.
Deshimaru
Shungo
S
Oda
Toshiro
T
Fujiwara
Satoru
S
D008841:Actin Cytoskeleton / Q000502:physiology*; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002417:Cattle; D009119:Muscle Contraction / Q000502:physiology; D018482:Muscle, Skeletal; D015879:Myosin Subfragments / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014336:Troponin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D015879:Myosin Subfragments; D011994:Recombinant Proteins; D014336:Troponin; D002118:Calcium