• BMB Reports
• /
• v.42 no.6
• /
• pp.350-355
• /
• 2009

Although previous studies on hexokinase (HK) II indicate both the N- and C-terminal halves are catalytically active, we show in this study the N-terminal half is significantly more catalytic than the C-terminal half in addition to having a significantly higher $K_m$ for ATP and Glu. Furthermore, truncated forms of intact HK II lacking its first N-terminal 18 amino acids ($\Delta$18) and a truncated N-terminal half lacking its first 18 amino acids ($\Delta$18N) have higher catalytic activity than other mutants tested. Similar results were obtained by PET-scan analysis using $^{18}F-FDG$. Our results collectively suggest that each domain of HK II possesses enzyme activity, unlike HK I, with the N-terminal half showing higher enzyme activity than the C-terminal half.

• The Korean Journal of Nuclear Medicine
• /
• v.33 no.1
• /
• pp.1-10
• /
• 1999

Cancer cells are known to show increased rates of glycolysis metabolism. Based on this, PET studies using F-18-fluorodeoxyglucose have been used for the detection of primary and metastatic tumors. To account for this increased glucose uptake, a variety of mechanisms has been proposed. Glucose influx across the cell membrane is mediated by a family of structurally related proteins known as glucose transporters (Gluts). Among 6 isoforms of Gluts, Glut-1 and/or Glut-3 have been reported to show increased expression in various tumors. Increased level of Glut mRNA transcription is supposed to be the basic mechanism of Glut overexpression at the protein level. Some oncogens such as src or ras intensely stimulate Glut-1 by means of increased Glut-1 mRNA levels. Hexokinase activity is another important factor in glucose uptake in cancer cells. Especially hexokinase type II is considered to be involved in glycolysis of cancer cells. Much of the hexokinase of tumor cells is bound to outer membrane of mitochondria by the porin, a hexokinase receptor. Through this interaction, hexokinase may gain preferred access to ATP synthesized via oxidative phosphorylation in the inner mitochondria compartment. Other biologic factors such as tumor blood flow, blood volume, hypoxia, and infiltrating cells in tumor tissue are involved. Relative hypoxia may activate the anaerobic glycotytic pathway. Surrounding macrophages and newly formed granulation tissue in tumor showed greater glucose uptake than did viable cancer cells. To expand the application of FDG PET in oncology, it is important for nuclear medicine physicians to understand the related mechanisms of glucose uptake in cancer tissue.

• Journal of Microbiology and Biotechnology
• /
• v.17 no.4
• /
• pp.638-643
• /
• 2007

The full encoding sequence for human type II hexokinase (HXK II) was cloned into the E. coli expression vector pET 21b and expressed as a C-terminally hexahistidine-tagged protein in the BL2l (DE3) strain. The IPTG-induced HXK II approximately accounted for 17% of the total E. coli proteins, and 81% of HXK $II_{6{\times}His}$ existed in inclusion bodies. To improve the production of soluble recombinant HXK II protein, in the functionally active form, we used low temperature, and the osmotic stress expression method. When expressed at $18^{\circ}C$, about 83% of HXK $II_{6{\times}His}$ existed in the soluble fraction, which amounted to a 4.1-fold yield over that expressed at $37^{\circ}C$. The soluble form of HXK $II_{6{\times}His}$ was also highly produced in the presence of 1M sorbitol under the standard condition $(37^{\circ}C)$, which indicated that temperature downshift and low water potentials were required to improve the yield of active recombinant HXK II protein. The expressed protein was purified by metal chelate affinity chromatography performed in an IDA Excellose column charged with $Ni^{2+}$ ions, resulting in about 40mg recombinant HXK II protein obtained with purity over 89% from 51 of E. coli culture. The identity of HXK $II_{6{\times}His}$ was confirmed by Western blotting analysis. Taken together, using the stress-governed expression described in this study, human active HXK II can be purified in sufficient amounts for biochemical and biomedical studies.

• 대한두경부종양학회:학술대회논문집
• /
• 2006.11a
• /
• pp.203-203
• /
• 2006

• Journal of Plant Biology
• /
• v.12 no.2
• /
• pp.15-22
• /
• 1969

By spectrophotometric assay method, the following enzymes could be detected in Dunaliella tertiolecta and Chlorella pyrenoidosa cell-free extracts: Hexokinase; Glucose-6-phosphate, 6-Phosphogluconate and Triosephosphate dehydrogenase; Transketolase; Phosphogluco and Ribosephosphate isomerase; Phosphoglucomutase; Phosphofructokinase; Fructosediphosphate aldorase and Ribulosephosphate 3-epimerase. Such enzymes are in accordance with the proposed pathway of glucose catabolism by D. tertiolecta as well as C. pyrenoidosa. Also, it could be estimated, under the presence of NADP, that pentose phosphate pathway were more active than glycolytic pathway in D. tertiolecta cell-free systems.

• Food Science and Preservation
• /
• v.7 no.1
• /
• pp.1-7
• /
• 2000

To develop natural antimicrobial agents for keeping qualities of postharvested greenhouse produce the antimiocrobial actions of Polygonum cuspidatum Sieb. et Zucc. extract , which showed remarkable antimicrobial effects against microorganism causing the postharvest decay of greenhouse produce, were investigate. In the inhibitory experiment of enzymes related to energy production metabolism hexokinase activities decreased to 73% and 68% by treating with Polygonum cuspidatum Sieb. et.Zucc. extract and Eugenia caryophyllata Thumnberg extract in comparison with control, respectively. Direct visualization of microbial cells by using both transmission electron microscope and scanning electron microscope showed that microbial cell membrane was destroyed by treating with the dilute extract solution. this change of celluloar membrane permeability could be identified in the experiment that 0-nitrophenyl-${\beta}$-D-galactopyrano-side(ONPG), the artificial substrate of ${\beta}$-galactosidase, was hydrolyzed in the presence of the extract, indicating that the membrane was perturbed. The separation and identification of the most antimicrobialo substances isolated from Polygonum cuspidatum Sieb et. Zucc. extract and Eugenia caryophyllata Thunberg extract were carried out by using gas chromatography and mass spectrometry 9GC/MSD), which were identified as eugenol. As a result, the functionality of Polygonum cuspidatum Sieb. et Zucc. extract and Eugenia caryophyllata Thunberg extract as antimicrobial agents for keeping qualities of postharvested greenhouse produce may be recommended.

• BMB Reports
• /
• v.50 no.12
• /
• pp.601-609
• /
• 2017

Mammalian target of rapamycin complex 1 (mTORC1) plays a major role in cell growth, proliferation, polarity, differentiation, development, and controls transitioning between anabolic and catabolic states of the cell. It collects almost all extracellular and intracellular signals from growth factors, nutrients, and maintains cellular homeostasis, and is involved in several pathological conditions including, neurodegeneration, Type 2 diabetes (T2D), obesity, and cancer. In this review, we summarize current knowledge of upstream signaling of mTORC1 to explain etiology of T2D and hypertriglyceridemia, in which state, the role of telomere attrition is explained. We discuss if chronic inhibition of mTORC1 can reverse adverse effects resulting from hyperactivation. In conclusion, we suggest the regulatory roles of telomerase (TERT) and hexokinase II (HKII) on mTORC1 as possible remedies to treat hyperactivation. The former inhibits mTORC1 under nutrientrich while the latter under starved condition. We provide an idea of TOS (TOR signaling) motifs that can be used for regulation of mTORC1.

• BMB Reports
• /
• v.51 no.1
• /
• pp.45-53
• /
• 2018

Mammalian target of rapamycin complex 1 (mTORC1) plays a major role in cell growth, proliferation, polarity, differentiation, development, and controls transitioning between anabolic and catabolic states of the cell. It collects almost all extracellular and intracellular signals from growth factors, nutrients, and maintains cellular homeostasis, and is involved in several pathological conditions including, neurodegeneration, Type 2 diabetes (T2D), obesity, and cancer. In this review, we summarize current knowledge of upstream signaling of mTORC1 to explain etiology of T2D and hypertriglyceridemia, in which state, the role of telomere attrition is explained. We discuss if chronic inhibition of mTORC1 can reverse adverse effects resulting from hyperactivation. In conclusion, we suggest the regulatory roles of telomerase (TERT) and hexokinase II (HKII) on mTORC1 as possible remedies to treat hyperactivation. The former inhibits mTORC1 under nutrient-rich while the latter under starved condition. We provide an idea of TOS (TOR signaling) motifs that can be used for regulation of mTORC1.

• Parasites, Hosts and Diseases
• /
• v.37 no.4
• /
• pp.229-236
• /
• 1999

In order to refer to the basic information regarding the identification of isolates obtained from a contact lens container in Korea, the isoelectric focusing gel electrophoresis was employed to compare the isoenzyme band patterns among Acanthamoeba spp. including eight isolates and the simple pairwise dissimilarity analysis was carried out. For an alkaline phosphate development, isolate 7 and Acanthamoeba polyphaga showed homologous band patterns, and isolates 1, 2, and 3 showed the same patterns. For lactate dehydrogenase, similar patterns were observed in isolates 2 and 3. Isolates 3 and 5 showed homologous band patterns for malate dehydrogenase and glucose phosphate isomerase. For hexokinase, isolates 4, 7, and A. hatchetti showed the same band patterns. In others, a considerable number of interstrain polymorphisms was observed in nine isoenzyme band patterns. In Acanthamoeba group II, genetic distances among isolates 1, 2, 3, 4, and 5 ranged from 0.104 to 0.200. In comparison to A. castellanii, A. hatchetti, and A. poIyphaga, genetic distances of isolates 7 and 8 were 0.254 and 0.219, respectively. In Acanthamoeba group III, including A. culbertsoni, A. healyi, and A. royreba, isolate 6 had genetic distances which ranged from 0.314 to 0.336. Finally, when comparing to the six reference Acanthamoeba, it was possible to classify isolates 1, 2, 3, 4, and 5, as genetically close-related species and as independent species group. Furthermore, isolates 6, 7 and 8 were identified as independent species as well.

• Asian-Australasian Journal of Animal Sciences
• /
• v.30 no.12
• /
• pp.1764-1772
• /
• 2017

Objective: This study investigated the effects of different dietary energy sources on early postmortem muscle metabolism of finishing pigs. Methods: Seventy-two barrow ($Duroc{\times}Landrace{\times}Yorkshire$, DLY) pigs ($65.0{\pm}2.0kg$) were allotted to three iso-energetic and iso-nitrogenous diets: A (44.1% starch, 5.9% crude fat, and 12.6% neutral detergent fibre [NDF]), B (37.6% starch, 9.5% crude fat, and 15.4% NDF) or C (30.9% starch, 14.3% crude fat, and 17.8% NDF). After the duration of 28-day feeding experiment, 24 pigs (eight per treatment) were slaughtered and the M. longissimus lumborum (LL) samples at 45 min postmortem were collected. Results: Compared with diet A, diet C resulted in greater adenosine triphosphate and decreased phosphocreatine (PCr) concentrations, greater activity of creatine kinase and reduced percentage bound activities of hexokinase (HK), and pyruvate kinase (PK) in LL muscles (p<0.05). Moreover, diet C decreased the phosphor-AKT level and increased the hydroxy-hypoxia-inducible $factor-1{\alpha}$ ($HIF-1{\alpha}$) level, as well as decreased the bound protein expressions of HK II, PKM2, and lactate dehydrogenase A (p<0.05). Conclusion: Diet C with the lowest level of starch and the highest levels of fat and NDF could enhance the PCr utilization and attenuate glycolysis early postmortem in LL muscle of finishing pigs.