title: Molecular and biochemical characterization of Eimeria tenella hexokinase.
abstract: Hexokinase (HK) is one of the key enzymes in the glycolytic pathway that catalyzes the phosphorylation of glucose. In the present study, we cloned the HK gene from the coccidian Eimeria tenella (EtHK), expressed EtHK as a His-tagged fusion protein, and characterized its primary biochemical features. Mutagenesis confirmed that residues S159, N216, and D217 are essential or important to the EtHK catalytic activity. EtHK exhibited high affinity for D-glucose (Km = 0.67 to 0.79 mM), but was also able to utilize 2-deoxy-D-glucose (Km = 5.66 mM), D-fructose (Km = 13.76 mM), and D-mannose (Km = 25.41 mM). We also observed that quercetin and mangiferin could inhibit the EtHK enzyme activity (IC50 values = 6.52 and 85.82 μM, respectively). Among the two inhibitors, mangiferin also inhibited the growth of E. tenella in vitro (MIC50 = 0.12 μM). These observations suggest that EtHK may be explored as potential drug target, and mangiferin and its analogs may be explored for developing anti-coccidial therapeutics.
authors.affiliation: Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, Guangdong Province, 510640 People's Republic of China.|Open Laboratory of Veterinary Public Health and Key Laboratory of Livestock Disease Prevention of Guangdong Province, Guangzhou, 510640, People's Republic of China.
authors.forename: Mingfei
authors.initials: M
authors.lastname: Sun
authors.affiliation: Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, Guangdong Province, 510640 People's Republic of China.|Open Laboratory of Veterinary Public Health and Key Laboratory of Livestock Disease Prevention of Guangdong Province, Guangzhou, 510640, People's Republic of China.
authors.forename: Shenquan
authors.initials: S
authors.lastname: Liao
authors.affiliation: College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou, Henan Province, 450002, People's Republic of China.
authors.forename: Longxian
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, Guangdong Province, 510640 People's Republic of China.|Open Laboratory of Veterinary Public Health and Key Laboratory of Livestock Disease Prevention of Guangdong Province, Guangzhou, 510640, People's Republic of China.
authors.forename: Caiyan
authors.initials: C
authors.lastname: Wu
authors.affiliation: Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, Guangdong Province, 510640 People's Republic of China.|Open Laboratory of Veterinary Public Health and Key Laboratory of Livestock Disease Prevention of Guangdong Province, Guangzhou, 510640, People's Republic of China.
authors.forename: Nanshan
authors.initials: N
authors.lastname: Qi
authors.affiliation: Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, Guangdong Province, 510640 People's Republic of China.|Open Laboratory of Veterinary Public Health and Key Laboratory of Livestock Disease Prevention of Guangdong Province, Guangzhou, 510640, People's Republic of China.
authors.forename: Minna
authors.initials: M
authors.lastname: Lv
authors.affiliation: Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, Guangdong Province, 510640 People's Republic of China.|Open Laboratory of Veterinary Public Health and Key Laboratory of Livestock Disease Prevention of Guangdong Province, Guangzhou, 510640, People's Republic of China.
authors.forename: Juan
authors.initials: J
authors.lastname: Li
authors.affiliation: Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, Guangdong Province, 510640 People's Republic of China.|Open Laboratory of Veterinary Public Health and Key Laboratory of Livestock Disease Prevention of Guangdong Province, Guangzhou, 510640, People's Republic of China.
authors.forename: Xuhui
authors.initials: X
authors.lastname: Lin
authors.affiliation: Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, Guangdong Province, 510640 People's Republic of China.|Open Laboratory of Veterinary Public Health and Key Laboratory of Livestock Disease Prevention of Guangdong Province, Guangzhou, 510640, People's Republic of China.
authors.forename: Jianfei
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, Guangdong Province, 510640 People's Republic of China.|Open Laboratory of Veterinary Public Health and Key Laboratory of Livestock Disease Prevention of Guangdong Province, Guangzhou, 510640, People's Republic of China.
authors.forename: Mingquan
authors.initials: M
authors.lastname: Xie
authors.affiliation: Department of Veterinary Pathobiology, College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, 77843-4467, Texas, USA. GZhu@cvm.tamu.edu.
authors.forename: Guan
authors.initials: G
authors.lastname: Zhu
authors.affiliation: Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, Guangdong Province, 510640 People's Republic of China. jianpicai@gmail.com.|Open Laboratory of Veterinary Public Health and Key Laboratory of Livestock Disease Prevention of Guangdong Province, Guangzhou, 510640, People's Republic of China. jianpicai@gmail.com.
authors.forename: Jianping
authors.initials: J
authors.lastname: Cai
mesh_terms: D003048:Coccidiosis / Q000469:parasitology; D016786:Eimeria tenella / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D004795:Enzyme Stability; D005632:Fructose / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006593:Hexokinase / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008358:Mannose / Q000378:metabolism; D010766:Phosphorylation; D015800:Protozoan Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism
chemical_list: D015800:Protozoan Proteins; D005632:Fructose; D006593:Hexokinase; D005947:Glucose; D008358:Mannose
keywords: Eimeria tenella; Glycolysis; Hexokinase; Mangiferin; Quercetin