Rapid relaxation of single frog skeletal muscle fibres following laser flash photolysis of the caged calcium chelator, diazo-2

AbstractDiazo-2 is a calcium chelator based on BAPTA [(1989) J. Biol. Chem., in press], whose electron withdrawing diazoacetyl group may be rapidly (2000 s−1) converted photochemically to an electron donating carboxymethyl group by exposure to near ultraviolet light, producing an increase in its calcium affinity (Kd changes from 2.2μM to 0.073 μM) without steric modification of the metal binding site. Photolysis of a 2 mM solution of this compound with a brief flash of light from a frequency-doubled ruby laser (347 nm) caused single skinned muscle fibres from the semitendinosus muscle of the frog Rana temporaria to relax with a mean half-time of 60.4±5 ms (range 30–100 ms, n = 15) at 12°C, which is faster than the relaxation observed in intact muscles (half-time 133 ms at 14°C [(1986) J. Mol. Biol. 188, 325–342]) and similar to the rate of the fast phase of tension decay in intact single fibres (20 s−1 at 1O°C [(1982) J. Physiol. 329, 1–20])