regulation and cytoprotective role of hexokinase iii监管和第三cytoprotective己糖激酶的作用.pdfVIP

Regulation and Cytoprotective Role of Hexokinase III 1 1 2 2,3 1 1 Eugene Wyatt , Rongxue Wu , Wael Rabeh , Hee-Won Park , Mohsen Ghanefar , Hossein Ardehali * 1 Division of Cardiology, Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States of America, 2 Structural Genomics Consortium, University of Toronto, Toronto, Canada, 3 Department of Pharmacology, University of Toronto, Toronto, Canada Abstract Background: Hexokinases (HKs) catalyze the first step in glucose metabolism. Of the three mammalian 100-kDa HK isoforms, HKI and II can bind to mitochondria and protect against cell death. HKIII does not bind mitochondria, and little is known about its regulation or cytoprotective effects. We studied the regulation of HKIII at the transcriptional and protein levels and investigated its role in cellular protection. Methodology/Principal Findings: We show that like HKII, HKIII expression is regulated by hypoxia, but other factors that regulate HKII expression have no effect on HKIII levels. This transcriptional regulation is partially dependent on hypoxia- inducible factor (HIF) signaling. We also demonstrate regulation at the protein level, as mutations in putative N-terminal substrate binding residues altered C-terminal catalytic activity, suggesting that HKIII activity is governed, in part, by interactions between these two domains. Overexpression of HKIII reduced oxidant-induced cell death, increased ATP levels, decreased the production of reactive oxygen species (ROS), and preserved mitochondrial membrane potential. HKIII overexpres