Disruption of Hexokinase II–Mitochondrial Binding Blocks Ischemic Preconditioning and Causes Rapid Cardiac Necrosis

• Published in: Circulation research Vol. 108; no. 10; pp. 1165 - 1169
• Main Authors: Smeele, Kirsten M.A, Southworth, Richard, Wu, Rongxue, Xie, Chaoqin, Nederlof, Rianne, Warley, Alice, Nelson, Jessica K, van Horssen, Pepijn, van den Wijngaard, Jeroen P, Heikkinen, Sami, Laakso, Markku, Koeman, Anneke, Siebes, Maria, Eerbeek, Otto, Akar, Fadi G, Ardehali, Hossein, Hollmann, Markus W, Zuurbier, Coert J
• Format: Journal Article
• Language: English
• Published: Hagerstown, MD American Heart Association, Inc 13.05.2011; Lippincott Williams & Wilkins
• Subjects: Animals; Biological and medical sciences; Fundamental and applied biological sciences. Psychology; Genetic Carrier Screening; Hexokinase - deficiency; Hexokinase - genetics; Hexokinase - metabolism; Ischemic Preconditioning, Myocardial - methods; Male; Membrane Potential, Mitochondrial - genetics; Mice; Mice, Inbred C57BL; Mice, Knockout; Mitochondria, Heart - enzymology; Mitochondria, Heart - genetics; Mitochondria, Heart - pathology; Myocardium - enzymology; Myocardium - pathology; Necrosis - enzymology; Necrosis - genetics; Necrosis - pathology; Protein Binding - genetics; Rats; tat Gene Products, Human Immunodeficiency Virus - physiology; Time Factors; Vertebrates: cardiovascular system; Vertebrata; Mitochondria; Mammalia; Reperfusion; Ischemia; Peptides; TAT peptide; Cardiovascular disease; Circulatory system; Preconditioning; Necrosis; cardiac physiology
• ISSN: 0009-7330; 1524-4571
• DOI: 10.1161/CIRCRESAHA.111.244962

RATIONALE:Isoforms I and II of the glycolytic enzyme hexokinase (HKI and HKII) are known to associate with mitochondria. It is unknown whether mitochondria-bound hexokinase is mandatory for ischemic preconditioning and normal functioning of the intact, beating heart. OBJECTIVE:We hypothesized that reducing mitochondrial hexokinase would abrogate ischemic preconditioning and disrupt myocardial function. METHODS AND RESULTS:Ex vivo perfused HKII hearts exhibited increased cell death after ischemia and reperfusion injury compared with wild-type hearts; however, ischemic preconditioning was unaffected. To investigate acute reductions in mitochondrial HKII levels, wild-type hearts were treated with a TAT control peptide or a TAT-HK peptide that contained the binding motif of HKII to mitochondria, thereby disrupting the mitochondrial HKII association. Mitochondrial hexokinase was determined by HKI and HKII immunogold labeling and electron microscopy analysis. Low-dose (200 nmol/L) TAT-HK treatment significantly decreased mitochondrial HKII levels without affecting baseline cardiac function but dramatically increased ischemia-reperfusion injury and prevented the protective effects of ischemic preconditioning. Treatment for 15 minutes with high-dose (10 μmol/L) TAT-HK resulted in acute mitochondrial depolarization, mitochondrial swelling, profound contractile impairment, and severe cardiac disintegration. The detrimental effects of TAT-HK treatment were mimicked by mitochondrial membrane depolarization after mild mitochondrial uncoupling that did not cause direct mitochondrial permeability transition opening. CONCLUSIONS:Acute low-dose dissociation of HKII from mitochondria in heart prevented ischemic preconditioning, whereas high-dose HKII dissociation caused cessation of cardiac contraction and tissue disruption, likely through an acute mitochondrial membrane depolarization mechanism. The results suggest that the association of HKII with mitochondria is essential for the protective effects of ischemic preconditioning and normal cardiac function through maintenance of mitochondrial potential.