title: A reevaluation of the roles of hexokinase I and II in the heart.
abstract: Hexokinase is responsible for glucose phosphorylation, a process fundamental to regulating glucose uptake. In some tissues, hexokinase translocates to the mitochondria, thereby increasing its efficiency and decreasing its susceptibility to product inhibition. It may also decrease free radical formation in the mitochondria and prevent apoptosis. Whether hexokinase translocation occurs in the heart is controversial; here, using immunogold labeling for the first time, we provide evidence for this process. Rat hearts (6 groups, n = 6/group), perfused with either glucose- or glucose + oleate (0.4 mmol/l)-containing buffer, were exposed to 30-min insulin stimulation, ischemia, or control perfusion. Hexokinase I (HK I) and hexokinase II (HK II) distributions were then determined. In glucose-perfused hearts, HK I-mitochondrial binding increased from 0.41 +/- 0.04 golds/mm in control hearts to 0.71 +/- 0.10 golds/mm after insulin and to 1.54 +/- 0.38 golds/mm after ischemia (P < 0.05). Similarly, HK II-mitochondrial binding increased from 0.16 +/- 0.02 to 0.53 +/- 0.08 golds/mm with insulin and 0.44 +/- 0.07 golds/mm after ischemia (P < 0.05). Under basal conditions, the fraction of HK I that was mitochondrial bound was five times greater than for HK II; insulin and ischemia caused a fourfold increase in HK II binding but only a doubling in HK I binding. Oleate decreased hexokinase-mitochondrial binding and abolished insulin-mediated translocation of HK I. Our data show that mitochondrial-hexokinase binding increases under insulin or ischemic stimulation and that this translocation is modified by oleate. These events are isoform specific, suggesting that HK I and HK II are independently regulated and implying that they perform different roles in cardiac glucose regulation.
authors.affiliation: The NMR Laboratory, Division of Imaging Sciences, 5th Floor Thomas Guy House, Guy's Hospital, St. Thomas' St., London SE1 9RT, UK. richard.southworth@kcl.ac.uk
authors.forename: Richard
authors.initials: R
authors.lastname: Southworth
authors.forename: Katherine A B
authors.initials: KA
authors.lastname: Davey
authors.forename: Alice
authors.initials: A
authors.lastname: Warley
authors.forename: Pamela B
authors.initials: PB
authors.lastname: Garlick
mesh_terms: D000818:Animals; D006321:Heart / Q000187:drug effects*; D006593:Hexokinase / Q000378:metabolism*; D007328:Insulin / Q000008:administration & dosage*; D008297:Male; D008929:Mitochondria, Heart / Q000187:drug effects / Q000378:metabolism*; D017202:Myocardial Ischemia / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism*; D020033:Protein Isoforms / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D014018:Tissue Distribution
chemical_list: D007328:Insulin; D020033:Protein Isoforms; D006593:Hexokinase