title: Both isoforms of ketohexokinase are dispensable for normal growth and development.
abstract: Dietary fructose intake has dramatically increased over recent decades and is implicated in the high rates of obesity, hypertension, and type 2 diabetes (metabolic syndrome) in Western societies. The molecular determinants of this epidemiologic correlation are incompletely defined, but high-flux fructose catabolism initiated by ketohexokinase (Khk, fructokinase) is believed to be important. The Khk gene encodes two enzyme isoforms with distinctive substrate preferences, the independent physiological roles of which are unclear. To investigate this question, and for testing the importance of Khk in metabolic syndrome, isoform-selective genetic lesions would be valuable. Two deficiency alleles of the mouse Khk gene were designed. The first, Khk(3a), uses targeted "knock-in" of a premature termination codon to induce a selective deficiency of the minor Khk-A isoform, preserving the major Khk-C isoform. The second, the Khk(Δ) allele, ablates both isoforms. Mice carrying each of these Khk-deficiency alleles were generated and validated at the DNA, RNA, and protein levels. Comparison between normal and knockout animals confirmed the specificity of the genetic lesions and allowed accurate analysis of the cellular distribution of Khk within tissues such as gut and liver. Both Khk(3a/3a) and Khk(Δ/Δ) homozygous mice were healthy and fertile and displayed minimal biochemical abnormalities under basal dietary conditions. These studies are the first demonstration that neither Khk isoform is required for normal growth and development. The new mouse models will allow direct testing of various hypotheses concerning the role of this enzyme in metabolic syndrome in humans and the value of Khk as a pharmacological target.
authors.affiliation: Leeds Institute of Molecular Medicine, University of Leeds, St James's University Hospital, United Kingdom.
authors.forename: C P
authors.initials: CP
authors.lastname: Diggle
authors.forename: M
authors.initials: M
authors.lastname: Shires
authors.forename: C
authors.initials: C
authors.lastname: McRae
authors.forename: D
authors.initials: D
authors.lastname: Crellin
authors.forename: J
authors.initials: J
authors.lastname: Fisher
authors.forename: I M
authors.initials: IM
authors.lastname: Carr
authors.forename: A F
authors.initials: AF
authors.lastname: Markham
authors.forename: B E
authors.initials: BE
authors.lastname: Hayward
authors.forename: A
authors.initials: A
authors.lastname: Asipu
authors.forename: D T
authors.initials: DT
authors.lastname: Bonthron
mesh_terms: D000818:Animals; D005260:Female; D005631:Fructokinases / Q000235:genetics* / Q000378:metabolism; D005632:Fructose; D015870:Gene Expression; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism
chemical_list: D020033:Protein Isoforms; D005632:Fructose; D005631:Fructokinases; C021475:ketohexokinase