Flash photolysis studies of relaxation and cross-bridge detachment: higher sensitivity of tonic than phasic smooth muscle to MgADP

Abstract

The effects of MgADP and inorganic phosphate (Pi) on cross-bridge detachment were determined in tonic (rabbit femoral artery) and phasic (rabbit bladder and guinea pig portal vein) smooth muscles permeabilized with staphylococcal alpha-toxin. Relaxation from rigor was induced by photolysis of ATP (1.2-1.5 mM) from caged ATP. The initial one second of relaxation from rigor was resolved into two exponential components: a rapid component with normalized amplitudes, Af, of 8, 15 and 26% and rate constants, kf (in s-1) of 26, 36 and 30 in rabbit femoral artery, guinea pig portal vein, and rabbit bladder; the respective rate constants of the second, slower component, ks, were 0.07, 0.2 and 0.1. Removal of residual endogenous ADP with apyrase treatment increased the amplitude Af and accelerated ks; addition of MgADP reduced Af. The combination of these effects (increases in Af and ks) decreased the t1/2 of relaxation from control values by factors of 2.6 (femoral artery), 6.7 (portal vein) and 10 (bladder). Pi (30 mM) further increased the amplitudes Af. The affinity of MgADP for myosin cross-bridges, estimated as the reduction of the relative amplitude of the rapid component, Af, was significantly higher in tonic than in phasic smooth muscle: the KD of MgADP was 1.1 +/- 0.3 microM in rabbit femoral artery and 4.9 +/- 1.0 microM in rabbit bladder. The higher affinity of tonic smooth muscle myosin for MgADP correlated with its relatively high LC17b isoform content (58 +/- 4.2%) in contrast to the lower affinity of the phasic, bladder detrusor smooth muscle that contained only the LC17a isoform.(ABSTRACT TRUNCATED AT 250 WORDS)