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By Freud Sigmund

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The reaction constant can be determined directly and the enzymic activity is linear dependent on the enzyme concentration. The yield of superoxide is extremely high and the method is highly sensitive. The distinction between catalytic and non-catalytic reactions is possible. For example, the reactivity of superoxide dismutase was found to be catalytically and of second order. 6 x 109 M - i s -1 at neutral pH are reported, thus being near the diffusion control 8s, 181,182,188-190). By way of contrast, caeruloplasmin the copper protein of the blood serum was demonstrated to react only stoichiometrically with superoxide 191) 31 Alfred G/irtner and Ulrich Weser Another advantage of pulse radiolysis lies in the evaluation of the catalytic scavenging of superoxide by low molecular mass complexes.

A recent overview is provided by Table 5. Direct Assays for Superoxide Dismutase Technique Specification EPR Rapid-freeze EPR 1~) 02 production with iron-sulfur proteins 17m Steady state EPR 177) Stopped flow spectrometry 178 Alkaline solution 179) 19F-NMR spectrometry 1so) Potassium superoxide in aprotic solvents NMR Simultaneous generation and determination of superoxide Electrochemical methods 30 Pulse radiolysis t81,182~ Flash photolysis 183) Opsonised graphite electrode 184~ Polarography ~85) Molecular and Functional Aspects of Superoxide Dismutases Asmus 186) This method is applicable for the determination of rate constants as well as exact measurements of the kinetic reaction orders.

Coli, A. vinelandii and P. 27 V 247). 0. 0, which is not observed above this pH-value. Both, iron-containing and mangano superoxide dismutases bear a net negative charge at physiological pH-values. The enzymes from E. coli are less active with increasing ionic strength 243). Acetylation of the lysine residues invert these effects. Therefore, a cationic group near the active site must be involved in the electrostatically facilitated catalysis. This observation parallels the ionic strength effects seen with Cu2Zn2superoxide dismutases (see Chapt.

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