To define the relation between the phosphoryl transfer via creatine kinase and the ability to recover from an ischemia-reperfusion challenge, the authors chemically inhibited creatine kinase activity with iodoacetamide (IAm) and then measured myocardial recovery after 2, 10, or 30 min of global ischemia followed by 30 min of reperfusion in the isolated, arterially perfused interventricular septa of the rabbit heart. During normoxia, IAm (0.5 M perfused for 15 min) did not by itself modify developed tension, maximal rate of tension development, or resting tension. In ischemia, IAm pretreatment increased the rate of developed tension loss and highly diminished developed tension recovery after reperfusion for all the ischemia periods tested. Moreover, IAm significantly enhanced the maximal increase in the resting tension induced by 10 or 30 min of ischemia plus reperfusion. Lactate dehydrogenase activity in reperfusion was also significantly increased over untreated septa. On the basis of the present results, the authors suggest that the aggravating effects exhibited by IAm on the ischemic myocardium are compatible with its creatine kinase inhibition properties and that creatine kinase activity is essential for full recovery from an ischemia-reperfusion challenge.