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The reaction of nitric oxide (NO) with metallothionein (MT) has been investigated at neutral pH under strictly anaerobic conditions. It is observed that NO mediates zinc release from MT by destroying zinc-sulphur clusters, but that it does not by itself S-nitrosylate MT in contrast to common belief. Zinc release and loss of thiolate groups under anaerobic conditions is found to be much slower than under aerobic conditions. The observed percentage loss of Zn(2+) and thiolate groups after 3 h of NO treatment are 62 and 39%, respectively. The reaction of NO with cysteine is reinvestigated and it is found that cysteine is quantitatively converted to cystine after 5 min of NO treatment at pH 7.3. At lower pH, a much lower rate of conversion is observed confirming the base-catalysed nature of the reaction of NO with thiols. On the basis of these results, a reaction mechanism involving electrophilic attack of NO on thiolate groups and subsequent formation of a nitrogen-centred radical, MTSN(. )OH, as intermediate is proposed for the reaction of NO with MT that leads to zinc release.

Nitric oxide induces Zn2+ release from metallothionein by destroying zinc–sulphur clusters without concomitant formation of S-nitrosothiol