Author:
- INTRODUCTION
- MATERIALS AND METHODS
- Structural-based modeling of recombinant PPDK and virtual screening of potential inhibitors.
- Preparation of stock solutions.
- Parasite and culture conditions.
- Expression and purification of recombinant protein.
- rPPDK enzyme assay.
- Bioactivity analysis of NCI compounds as potential inhibitors against rPPDK.
- In vitro screening of selected compounds against E. histolytica trophozoites by a minimum inhibitory concentration (MIC) assay.
- In vitro cell viability assay to determine IC50 value.
- RESULTS
- DISCUSSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
Stanley SL Jr, 2006. Vaccines for amoebiasis: barriers and opportunities. Parasitol 133: 81–86.
-
2.
Salles JM, Salles MJ, Moraes LA, Silva MC, 2007. Invasive amebiasis: an update on diagnosis and management. Expert Rev Anti Infect Ther 5: 893–901.
-
3.
Fotedar R, Stark D, Beebe N, Marriott D, Ellis J, Harkness J, 2007. Laboratory diagnostic techniques for Entamoeba species. Clin Microbiol Rev 20: 511–532.
-
4.
Baxt LA, Singh U, 2008. New insights into Entamoeba histolytica pathogenesis. Curr Opin Infect Dis 21: 489–494.
-
5.
Behnia M, Haghighi A, Komeylizadeh H, Tabaei SJS, Abadi A, 2008. Inhibitory effects of Iranian Thymus vulgaris extracts on in vitro growth of Entamoeba histolytica. Korean J Parasitol 46: 153–156.
-
6.
Upcroft P, Upcroft JA, 2001. Drug targets and mechanisms of resistance in the anaerobic protozoa. Clin Microbiol Rev 14: 150–164.
-
7.
Burchard GD, Mirelman D, 1988. Entamoeba histolytica: virulence potential and sensitivity to metronidazole and emetine of four isolates possessing non-pathogeniczymodemes. Exp Parasitol 66: 231–242.
-
8.
Samarawickrema NA, Brown DM, Upcroft JA, Thammapalerd N, Upcroft P, 1997. Involvement of superoxide dismutase and pyruvate: ferredoxin oxidoreductase in mechanisms of metronidazole resistance in Entamoeba histolytica. J Antimicrob Chemother 40: 833–840.
-
9.
Wassmann C, Hellberg A, Tannich A, Bruchhaus I, 1999. Metronidazole resistance in the protozoan parasite Entamoeba histolytica is associated with increased expression of iron-containing superoxide dismutase and peroxiredoxin and decreased expression of ferredoxin 1 and flavin reductase. J Biol Chem 274: 26051–26056.
-
10.
Adagu IS, Nolder D, Warhurst DC, Rossignol JF, 2002. In vitro activity of nitazoxanide and related compounds against isolates of Giardia intestinalis, Entamoeba histolytica and Trichomonas vaginalis. Antimicrob Agents Chemother 49: 103–111.
-
11.
Bansal D, Sehgal R, Chawla Y, Mahajan RC, Malla N, 2004. In vitro activity of antiamoebic drugs against clinical isolates of Entamoeba histolytica and Entamoeba dispar. Ann Clin Microbiol Antimicrob 3: 27.
-
12.
Huang B, Vetting MW, Roderick SL, 2005. The active site of O-acetylserine sulfhydrylase is the anchor point for bienzyme complex formation with serine acetyltransferase. J Bacteriol 187: 3201–3205.
-
13.
Saavedra E, Encalada R, Pineda E, Jasso-Chávez R, Moreno-Sánchez R, 2005. Glycolysis in Entamoeba histolytica. Biochemical characterization of recombinant glycolytic enzymes and flux control analysis. FEBS J 272: 1767–1783.
-
14.
Pineda E, Encalada R, Vázquez C, González Z, Moreno-Sánchez R, Saavedra E, 2015. Glucose metabolism and its controlling mechanisms in Entamoeba histolytica. Nozaki T, Battacharya A, eds. Amebiasis. Tokyo, Japan: Springer, 351–372.
-
15.
Saavedra E, Olivos A, Encalada R, Moreno-Sanchez R, 2004. Entamoeba histolytica: kinetic and molecular evidence of a previously unidentified pyruvate kinase. Exp Parasitol 106: 11–21.
-
16.
Bakker BM, Westerhoff HV, Opperdoes FR, Michels PA, 2000. Metabolic control analysis of glycolysis in trypanosomes as an approach to improve selectivity and effectiveness of drugs. Mol Biochem Parasitol 106: 1–10.
-
17.
Verlinde CL, Hannaert V, Blonski C, Willson M, Périé JJ, Fothergill-Gilmore LA, Opperdoes FR, Gelb MH, Hol WG, Michels PA, 2001. Glycolysis as a target for the design of new anti-trypanosome drugs. Drug Resist Updat 4: 50–65.
-
18.
Mansour TE, 2002. Chemotherapeutic Targets in Parasites: Contemporary Strategies. Cambridge, United Kingdom: Cambridge University Press.
-
19.
Eubank WB, Reeves RE, 1982. Analog inhibitors for the pyrophosphate-dependent phosphofructokinase of Entamoeba histolytica and their effect on culture growth. J Parasitol 1: 599–602.
-
20.
Bruchhaus I, Jacobs T, Denart M, Tannich E, 1996. Pyrophosphate dependent phosphofructokinase of Entamoeba histolytica: molecular cloning, recombinant expression and inhibition by pyrophosphate analogues. Biochem J 316: 57–63.
-
21.
Ye D et al.., 2001. Investigation of the catalytic site within the ATP-grasp domain of Clostridium symbiosum pyruvate phosphate dikinase. J Biol Chem 27: 37630–37639.
-
22.
Saidin S, Yunus MH, Zakaria ND, Razak KA, Huat LB, Othman N, Noordin R, 2014. Production of recombinant Entamoeba histolytica pyruvate phosphate dikinase and its application in a lateral flow dipstick test for amoebic liver abscess. BMC Infect Dis 14: 182.
-
23.
Saavedra-Lira E, Ramirez-Silva L, Perez-Montfort R, 1998. Expression and characterization of recombinant pyruvate phosphate dikinase from Entamoeba histolytica. Biochim Biophys Acta 1382: 47–54.
-
24.
Swami B, Lavakusuiu D, Sitha Devi C, 1977. Imidazole and metronidazole in the treatment of intestinal amoebiasis. Curr Med Res Opin 5: 152–156.
-
25.
Nagpal I, Raj I, Subbarao N, Gourinath S, 2012. Virtual screening, identification and fhrymn testing of novel inhibitors of O-acetyl-L-serine sulfhydrylase of Entamoeba histolytica. PLoS One 7: e30305.
-
26.
Becker S, Hoffman P, Houpt ER, 2011. Efficacy of antiamebic drugs in a mouse model. Am J Trop Med Hyg 84: 581–586.
-
27.
Hwang EW, Cheung L, Mojtahed A, Cartwright CA, 2011. Relapse of intestinal and hepatic amebiasis after treatment. Dig Dis Sci 56: 677–680.
-
28.
Cooper RA, Kornberg HL, 1974. Phosphoenolpyruvate synthetase and pyruvate, phosphate dikinase. Boyer PD, ed. The Enzymes. New York, NY: Academic Press, 631–649.
-
29.
Bringaud F, Baltz D, Baltz T, 1998. Functional and molecular characterization of a glycosomal PPi-dependent enzyme in trypanosomatids: pyruvate, phosphate dikinase. Proc Natl Acad Sci USA 95: 7963–7968.
-
30.
Savjani KT, Gajjar AK, Savjani JK, 2012. Drug solubility: importance and enhancement techniques. ISRN Pharm 2012: 195727.
-
31.
Khadka P et al.., 2014. Pharmaceutical particle technologies: an approach to improve drug solubility, dissolution and bioavailability. Asian J Pharmacol 9: 304–316.
-
32.
Kerns EH, Di L, 2003. Pharmaceutical profiling in drug discovery. Drug Discov Today 8: 316–323.
-
33.
Li D, Edward HK, 2015. Effects of properties on biological assays. Drug-Like Properties: Concepts, Structure Design and Methods from ADME to Toxicity Optimization. Cambridge, MA: Academic Press, 487–496.
-
34.
Agarwal SM, Jain R, Bhattacharya A, Azam A, 2008. Inhibitors of Escherichia coli serine acetyltransferase block proliferation of Entamoeba histolytica trophozoites. Int J Parasitol 38: 137–141.
-
35.
Stephen P, Vijayan R, Bhat A, Subbarao N, Bamezai RNK, 2008. Molecular modeling on pyruvate phosphate dikinase of Entamoeba histolytica and in silico virtual screening for novel inhibitors. J Comput Aided Mol Des 22: 647–660.
-
36.
Saucedo-Mendiola ML, Salas-Pacheco JM, Nájera H, Rojo-Domínguez A, Yépez-Mulia L, Avitia-Domínguez C, Téllez-Valencia A, 2014. Discovery of Entamoeba histolytica hexokinase 1 inhibitors through homology modeling and virtual screening. J Enzyme Inhib Med Chem 29: 325–332.
-
37.
Pandey S, Kekre N, Naderi J, McNulty J, 2005. Induction of apoptotic cell death specifically in rat and human cancer cells by pancratistatin. Artif Cell Blood Sub 33: 279–295.
-
38.
El-Sayed NM, Ismail KA, Ahmed SAEG, Hetta MH, 2012. In vitro amoebicidal activity of ethanol extracts of Arachis hypogaea L., Curcuma longa L. and Pancratium maritimum L. on Acanthamoeba castellanii cysts. Parasitol Res 110: 1985–1992.
-
39.
Ouarzane-Amara M, Franetich JF, Mazier D, Pettit GR, Meijer L, Doerig C, Desportes-Livage I, 2001. In vitro activities of two antimitotic compounds, pancratistatin and 7-deoxynarciclasine, against Encephalitozoon intestinalis, a microsporidium causing infections in humans. Antimicrob Agents Chemother 45: 3409–3415.
-
40.
Gabrielsen B et al.., 1992. Antiviral (RNA) activity of selected Amaryllidaceae isoquinoline constituents and synthesis of related substances. J Nat Prod 55: 1569–1581.
-
41.
Reeves RE, 1968. A new enzyme with the glycolytic function of pyruvate kinase. J Biochem 243: 3202–3204.
-
42.
Hrdy I, Mertens E, Nohynkova E, 1993. Giardia intestinalis: detection and characterization of a pyruvate phosphate dikinase. Exp Parasitol 76: 438–441.
-
43.
South DJ, Reeves RE, 1975. Pyruvate, orthophosphate dikinase from Bacteroides symbiosus. Methods Enzymol 42: 181–187.
-
44.
Edwards GE, Nakamoto H, Burnell JN, Hatch MD, 1985. Pyruvate, Pi dikinase and NADP-malate dehydrogenase in C4 photosynthesis: properties and mechanism of light/dark regulation. Annu Rev Plant Physiol 36: 255–286.
-
45.
Tjaden B, Plagens A, Dörr C, Siebers B, Hensel R, 2006. Phosphoenolpyruvate synthetase and pyruvate, phosphate dikinase of Thermoproteus tenax: key pieces in the puzzle of archaeal carbohydrate metabolism. Mol Microbiol 60: 287–298.
-
46.
Eisaki N, Tatsumi H, Murakami S, Horiuchi T, 1999. Pyruvate phosphate dikinase from a thermophilic actinomyces Microbispora rosea subsp. aerata: purification, characterization and molecular cloning of the gene. Biochim. Biophys Acta 1431: 363–373.
-
47.
Hiltpold A, Thomas RM, Köhler P, 1999. Purification and characterization of recombinant pyruvate phosphate dikinase from Giardia. Mol Biochem Parasitol 104: 157–169.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 13 | 13 | 5 |
| Full Text Views | 434 | 91 | 0 |
| PDF Downloads | 169 | 38 | 0 |
In Vitro Testing of Potential Entamoeba histolytica Pyruvate Phosphate Dikinase Inhibitors
Syazwan Saidin
Syazwan Saidin
Institute for Research in Molecular Medicine, Universiti Sains Malaysia, USM, Penang, Malaysia
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Nurulhasanah Othman
Nurulhasanah Othman
Institute for Research in Molecular Medicine, Universiti Sains Malaysia, USM, Penang, Malaysia
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Rahmah Noordin
Rahmah Noordin
Institute for Research in Molecular Medicine, Universiti Sains Malaysia, USM, Penang, Malaysia
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Adverse effects and resistance to metronidazole have motivated the search for new antiamoebic agents against Entamoeba histolytica. Control of amoeba growth may be achieved by inhibiting the function of the glycolytic enzyme and pyruvate phosphate dikinase (PPDK). In this study, we screened 10 compounds using an in vitro PPDK enzyme assay. These compounds were selected from a virtual screening of compounds in the National Cancer Institute database. The antiamoebic activity of the selected compounds was also evaluated by determining minimal inhibitory concentrations (MICs) and IC50 values using the nitro-blue tetrazolium reduction assay. Seven of the 10 compounds showed inhibitory activities against the adenosine triphosphate (ATP)/inorganic phosphate binding site of the ATP-grasp domain. Two compounds, NSC349156 (pancratistatin) and NSC228137 (7-ethoxy-4-[4-methylphenyl] sulfonyl-3-oxido-2, 1, 3-benzoxadiazol-3-ium), exhibited inhibitory effects on the growth of E. histolytica trophozoites with MIC values of 25 and 50 μM, and IC50 values of 14 and 20.7 μM, respectively.
Author Notes
Financial support: This work was supported by Malaysian Ministry of Higher Education (MOHE) FRGS Grant No. 203/CIPPM /6711241.
Authors’ addresses: Syazwan Saidin, Nurulhasanah Othman, and Rahmah Noordin, Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Penang, Malaysia, E-mails: syazwan_saidin@yahoo.com, nho80@yahoo.co.uk, and rahmah8485@gmail.com.
Reprint requests: Rahmah Noordin, Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800 USM, Penang, Malaysia, E-mails: rahmah8485@gmail.com or rahmah@usm.my.
- INTRODUCTION
- MATERIALS AND METHODS
- Structural-based modeling of recombinant PPDK and virtual screening of potential inhibitors.
- Preparation of stock solutions.
- Parasite and culture conditions.
- Expression and purification of recombinant protein.
- rPPDK enzyme assay.
- Bioactivity analysis of NCI compounds as potential inhibitors against rPPDK.
- In vitro screening of selected compounds against E. histolytica trophozoites by a minimum inhibitory concentration (MIC) assay.
- In vitro cell viability assay to determine IC50 value.
- RESULTS
- DISCUSSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
Stanley SL Jr, 2006. Vaccines for amoebiasis: barriers and opportunities. Parasitol 133: 81–86.
-
2.
Salles JM, Salles MJ, Moraes LA, Silva MC, 2007. Invasive amebiasis: an update on diagnosis and management. Expert Rev Anti Infect Ther 5: 893–901.
-
3.
Fotedar R, Stark D, Beebe N, Marriott D, Ellis J, Harkness J, 2007. Laboratory diagnostic techniques for Entamoeba species. Clin Microbiol Rev 20: 511–532.
-
4.
Baxt LA, Singh U, 2008. New insights into Entamoeba histolytica pathogenesis. Curr Opin Infect Dis 21: 489–494.
-
5.
Behnia M, Haghighi A, Komeylizadeh H, Tabaei SJS, Abadi A, 2008. Inhibitory effects of Iranian Thymus vulgaris extracts on in vitro growth of Entamoeba histolytica. Korean J Parasitol 46: 153–156.
-
6.
Upcroft P, Upcroft JA, 2001. Drug targets and mechanisms of resistance in the anaerobic protozoa. Clin Microbiol Rev 14: 150–164.
-
7.
Burchard GD, Mirelman D, 1988. Entamoeba histolytica: virulence potential and sensitivity to metronidazole and emetine of four isolates possessing non-pathogeniczymodemes. Exp Parasitol 66: 231–242.
-
8.
Samarawickrema NA, Brown DM, Upcroft JA, Thammapalerd N, Upcroft P, 1997. Involvement of superoxide dismutase and pyruvate: ferredoxin oxidoreductase in mechanisms of metronidazole resistance in Entamoeba histolytica. J Antimicrob Chemother 40: 833–840.
-
9.
Wassmann C, Hellberg A, Tannich A, Bruchhaus I, 1999. Metronidazole resistance in the protozoan parasite Entamoeba histolytica is associated with increased expression of iron-containing superoxide dismutase and peroxiredoxin and decreased expression of ferredoxin 1 and flavin reductase. J Biol Chem 274: 26051–26056.
-
10.
Adagu IS, Nolder D, Warhurst DC, Rossignol JF, 2002. In vitro activity of nitazoxanide and related compounds against isolates of Giardia intestinalis, Entamoeba histolytica and Trichomonas vaginalis. Antimicrob Agents Chemother 49: 103–111.
-
11.
Bansal D, Sehgal R, Chawla Y, Mahajan RC, Malla N, 2004. In vitro activity of antiamoebic drugs against clinical isolates of Entamoeba histolytica and Entamoeba dispar. Ann Clin Microbiol Antimicrob 3: 27.
-
12.
Huang B, Vetting MW, Roderick SL, 2005. The active site of O-acetylserine sulfhydrylase is the anchor point for bienzyme complex formation with serine acetyltransferase. J Bacteriol 187: 3201–3205.
-
13.
Saavedra E, Encalada R, Pineda E, Jasso-Chávez R, Moreno-Sánchez R, 2005. Glycolysis in Entamoeba histolytica. Biochemical characterization of recombinant glycolytic enzymes and flux control analysis. FEBS J 272: 1767–1783.
-
14.
Pineda E, Encalada R, Vázquez C, González Z, Moreno-Sánchez R, Saavedra E, 2015. Glucose metabolism and its controlling mechanisms in Entamoeba histolytica. Nozaki T, Battacharya A, eds. Amebiasis. Tokyo, Japan: Springer, 351–372.
-
15.
Saavedra E, Olivos A, Encalada R, Moreno-Sanchez R, 2004. Entamoeba histolytica: kinetic and molecular evidence of a previously unidentified pyruvate kinase. Exp Parasitol 106: 11–21.
-
16.
Bakker BM, Westerhoff HV, Opperdoes FR, Michels PA, 2000. Metabolic control analysis of glycolysis in trypanosomes as an approach to improve selectivity and effectiveness of drugs. Mol Biochem Parasitol 106: 1–10.
-
17.
Verlinde CL, Hannaert V, Blonski C, Willson M, Périé JJ, Fothergill-Gilmore LA, Opperdoes FR, Gelb MH, Hol WG, Michels PA, 2001. Glycolysis as a target for the design of new anti-trypanosome drugs. Drug Resist Updat 4: 50–65.
-
18.
Mansour TE, 2002. Chemotherapeutic Targets in Parasites: Contemporary Strategies. Cambridge, United Kingdom: Cambridge University Press.
-
19.
Eubank WB, Reeves RE, 1982. Analog inhibitors for the pyrophosphate-dependent phosphofructokinase of Entamoeba histolytica and their effect on culture growth. J Parasitol 1: 599–602.
-
20.
Bruchhaus I, Jacobs T, Denart M, Tannich E, 1996. Pyrophosphate dependent phosphofructokinase of Entamoeba histolytica: molecular cloning, recombinant expression and inhibition by pyrophosphate analogues. Biochem J 316: 57–63.
-
21.
Ye D et al.., 2001. Investigation of the catalytic site within the ATP-grasp domain of Clostridium symbiosum pyruvate phosphate dikinase. J Biol Chem 27: 37630–37639.
-
22.
Saidin S, Yunus MH, Zakaria ND, Razak KA, Huat LB, Othman N, Noordin R, 2014. Production of recombinant Entamoeba histolytica pyruvate phosphate dikinase and its application in a lateral flow dipstick test for amoebic liver abscess. BMC Infect Dis 14: 182.
-
23.
Saavedra-Lira E, Ramirez-Silva L, Perez-Montfort R, 1998. Expression and characterization of recombinant pyruvate phosphate dikinase from Entamoeba histolytica. Biochim Biophys Acta 1382: 47–54.
-
24.
Swami B, Lavakusuiu D, Sitha Devi C, 1977. Imidazole and metronidazole in the treatment of intestinal amoebiasis. Curr Med Res Opin 5: 152–156.
-
25.
Nagpal I, Raj I, Subbarao N, Gourinath S, 2012. Virtual screening, identification and fhrymn testing of novel inhibitors of O-acetyl-L-serine sulfhydrylase of Entamoeba histolytica. PLoS One 7: e30305.
-
26.
Becker S, Hoffman P, Houpt ER, 2011. Efficacy of antiamebic drugs in a mouse model. Am J Trop Med Hyg 84: 581–586.
-
27.
Hwang EW, Cheung L, Mojtahed A, Cartwright CA, 2011. Relapse of intestinal and hepatic amebiasis after treatment. Dig Dis Sci 56: 677–680.
-
28.
Cooper RA, Kornberg HL, 1974. Phosphoenolpyruvate synthetase and pyruvate, phosphate dikinase. Boyer PD, ed. The Enzymes. New York, NY: Academic Press, 631–649.
-
29.
Bringaud F, Baltz D, Baltz T, 1998. Functional and molecular characterization of a glycosomal PPi-dependent enzyme in trypanosomatids: pyruvate, phosphate dikinase. Proc Natl Acad Sci USA 95: 7963–7968.
-
30.
Savjani KT, Gajjar AK, Savjani JK, 2012. Drug solubility: importance and enhancement techniques. ISRN Pharm 2012: 195727.
-
31.
Khadka P et al.., 2014. Pharmaceutical particle technologies: an approach to improve drug solubility, dissolution and bioavailability. Asian J Pharmacol 9: 304–316.
-
32.
Kerns EH, Di L, 2003. Pharmaceutical profiling in drug discovery. Drug Discov Today 8: 316–323.
-
33.
Li D, Edward HK, 2015. Effects of properties on biological assays. Drug-Like Properties: Concepts, Structure Design and Methods from ADME to Toxicity Optimization. Cambridge, MA: Academic Press, 487–496.
-
34.
Agarwal SM, Jain R, Bhattacharya A, Azam A, 2008. Inhibitors of Escherichia coli serine acetyltransferase block proliferation of Entamoeba histolytica trophozoites. Int J Parasitol 38: 137–141.
-
35.
Stephen P, Vijayan R, Bhat A, Subbarao N, Bamezai RNK, 2008. Molecular modeling on pyruvate phosphate dikinase of Entamoeba histolytica and in silico virtual screening for novel inhibitors. J Comput Aided Mol Des 22: 647–660.
-
36.
Saucedo-Mendiola ML, Salas-Pacheco JM, Nájera H, Rojo-Domínguez A, Yépez-Mulia L, Avitia-Domínguez C, Téllez-Valencia A, 2014. Discovery of Entamoeba histolytica hexokinase 1 inhibitors through homology modeling and virtual screening. J Enzyme Inhib Med Chem 29: 325–332.
-
37.
Pandey S, Kekre N, Naderi J, McNulty J, 2005. Induction of apoptotic cell death specifically in rat and human cancer cells by pancratistatin. Artif Cell Blood Sub 33: 279–295.
-
38.
El-Sayed NM, Ismail KA, Ahmed SAEG, Hetta MH, 2012. In vitro amoebicidal activity of ethanol extracts of Arachis hypogaea L., Curcuma longa L. and Pancratium maritimum L. on Acanthamoeba castellanii cysts. Parasitol Res 110: 1985–1992.
-
39.
Ouarzane-Amara M, Franetich JF, Mazier D, Pettit GR, Meijer L, Doerig C, Desportes-Livage I, 2001. In vitro activities of two antimitotic compounds, pancratistatin and 7-deoxynarciclasine, against Encephalitozoon intestinalis, a microsporidium causing infections in humans. Antimicrob Agents Chemother 45: 3409–3415.
-
40.
Gabrielsen B et al.., 1992. Antiviral (RNA) activity of selected Amaryllidaceae isoquinoline constituents and synthesis of related substances. J Nat Prod 55: 1569–1581.
-
41.
Reeves RE, 1968. A new enzyme with the glycolytic function of pyruvate kinase. J Biochem 243: 3202–3204.
-
42.
Hrdy I, Mertens E, Nohynkova E, 1993. Giardia intestinalis: detection and characterization of a pyruvate phosphate dikinase. Exp Parasitol 76: 438–441.
-
43.
South DJ, Reeves RE, 1975. Pyruvate, orthophosphate dikinase from Bacteroides symbiosus. Methods Enzymol 42: 181–187.
-
44.
Edwards GE, Nakamoto H, Burnell JN, Hatch MD, 1985. Pyruvate, Pi dikinase and NADP-malate dehydrogenase in C4 photosynthesis: properties and mechanism of light/dark regulation. Annu Rev Plant Physiol 36: 255–286.
-
45.
Tjaden B, Plagens A, Dörr C, Siebers B, Hensel R, 2006. Phosphoenolpyruvate synthetase and pyruvate, phosphate dikinase of Thermoproteus tenax: key pieces in the puzzle of archaeal carbohydrate metabolism. Mol Microbiol 60: 287–298.
-
46.
Eisaki N, Tatsumi H, Murakami S, Horiuchi T, 1999. Pyruvate phosphate dikinase from a thermophilic actinomyces Microbispora rosea subsp. aerata: purification, characterization and molecular cloning of the gene. Biochim. Biophys Acta 1431: 363–373.
-
47.
Hiltpold A, Thomas RM, Köhler P, 1999. Purification and characterization of recombinant pyruvate phosphate dikinase from Giardia. Mol Biochem Parasitol 104: 157–169.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 13 | 13 | 5 |
| Full Text Views | 434 | 91 | 0 |
| PDF Downloads | 169 | 38 | 0 |