Physiological Studies of Oxygen Protection Mechanisms in the Heterocysts of Anabaena cylindrica

Abstract

The mechanism of O(2) protection of nitrogenase in the heterocysts of Anabaena cylindrica was studied in vivo. Resistance to O(2) inhibition of nitrogenase activity correlated with the O(2) tension of the medium in which heterocyst formation was induced. O(2) resistance also correlated with the apparent K(m) for acetylene, indicating that O(2) tension may influence the development of a gas diffusion barrier in the heterocysts. The role of respiratory activity in protecting nitrogenase from O(2) that diffuses into the heterocyst was studied using inhibitors of carbon metabolism. Reductant limitation induced by 3-(3,4-dichlorophenyl)-1, 1-dimethylurea increased the O(2) sensitivity of in vivo acetylene reduction. Azide, at concentrations (30 mM) sufficient to completely inhibit dark nitrogenase activity (a process dependent on oxidative phosphorylation for its ATP supply), severely inhibited short-term light-dependent acetylene reduction in the presence of O(2) but not in its absence. After 3 h of aerobic incubation in the presence of 20 mM azide, 75% of cross-reactive component I (Fe-Mo protein) in nitrogenase was lost; less than 35% was lost under microaerophilic conditions. Sodium malonate and monofluoroacetate, inhibitors of Krebs cycle activity, had only small inhibitory effects on nitrogenase activity in the light and on cross-reactive material. The results suggest that oxygen protection is dependent on both an O(2) diffusion barrier and active respiration by the heterocyst.