Comparative Analysis of the Interaction of Silver Nanoparticles with Hexokinase from Trypanosoma brucei and Humans
Madalitso M Mlozen,1,2 Jacqueline Van Marwijk,1 Brendan Shane Wilhelmi,1 Chris Whiteley1 1Department of Biochemistry and Microbiology, Rhodes University, Makhanda (Grahamstown), South Africa; 2Malawi Adventist University, Malamulo Campus, Department of Biomedical Sciences, Makwasa, MalawiCorresponde...
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Dove Medical Press
2023-03-01
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| Series: | International Journal of Nanomedicine |
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| Online Access: | https://www.dovepress.com/comparative-analysis-of-the-interaction-of-silver-nanoparticles-with-h-peer-reviewed-fulltext-article-IJN |
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| author | Mlozen MM Van Marwijk J Wilhelmi BS Whiteley C |
| author_facet | Mlozen MM Van Marwijk J Wilhelmi BS Whiteley C |
| author_sort | Mlozen MM |
| collection | DOAJ |
| description | Madalitso M Mlozen,1,2 Jacqueline Van Marwijk,1 Brendan Shane Wilhelmi,1 Chris Whiteley1 1Department of Biochemistry and Microbiology, Rhodes University, Makhanda (Grahamstown), South Africa; 2Malawi Adventist University, Malamulo Campus, Department of Biomedical Sciences, Makwasa, MalawiCorrespondence: Madalitso M Mlozen, Malawi Adventist University, Malamulo campus, Department of Biomedical Sciences, P.O.Box 55, Makwasa, Tel +265 884628334, Email mlozenim@mchs.advenist.orgBackground: Regardless of the efforts to ease cases of human African trypanosomiasis (HAT), an increased number of cases get reported annually. This is because of drug resistant Trypanosoma brucei (Tb), the causative agent of the illness. This has renewed the need for creative methods to find new anti-trypanosomal drugs. The blood stream form (BSF) of the parasite depends exclusively on the glycolytic pathway for energy production while it is in the human host. Interruptions in this pathway efficiently kills the parasite. Trypanosoma brucei hexokinase (TbHK) is the first enzyme in glycolysis, and any effectors or inhibitors of TbHK would have potential as anti-trypanosomal agents.Methods: TbHK and human glucokinase (hGCK) were over-expressed with a 6 histidine-tag in E. coli BL21(DE3) cells having the pRARE2 plasmid.Results: TbHK had thermal and pH stability between 30°C and 55°C and 7.5 and 8.5, respectively, while hGCK exhibited thermal and pH stability between 30°C and 40°C and 7.0 and 8.0, respectively. Kinetically, TbHK had a Km of 39.3 μM, Vmax of 0.066 μmol.min− 1.mL− 1, kcat of 2.05 min− 1 and kcat/Km of 0.0526 min− 1.μmol− 1. hGCK exhibited a Km of 4.5 μM, Vmax of 0.032 μnmol.min− 1.mL− 1, kcat of 11.25 min− 1, and kcat/Km of 2.5 min− 1.μmol− 1. Interaction kinetic studies of silver nanoparticles (AgNPs) (0.1 μM) of average size of 6 nm with TbHK and hGCK were conducted. AgNPs selectively inhibited TbHK over hGCK. TbHK showed a non-competitive inhibition with a 50% and 28% decrease in Vmax, and kcat/km, respectively. HsGCK showed a 33% increase in affinity, 9% decrease in Vmax, and a 50% increase in enzyme efficiency.Conclusion: The observed pattern of hGCK and AgNPs falls under the uncompetitive inhibition. The observed highly selective inhibitory effects of AgNPs between TbHK and hGCK may be used in development of new anti-trypanosomal drugs.Keywords: nanoparticles, hexokinase, trypanosomiasis, nanomedicine |
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| format | Article |
| id | doaj.art-38252105a18b4037ac320703e14714b5 |
| institution | Directory Open Access Journal |
| issn | 1178-2013 |
| language | English |
| last_indexed | 2024-04-09T22:00:38Z |
| publishDate | 2023-03-01 |
| publisher | Dove Medical Press |
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| series | International Journal of Nanomedicine |
| spelling | doaj.art-38252105a18b4037ac320703e14714b52023-03-23T18:06:52ZengDove Medical PressInternational Journal of Nanomedicine1178-20132023-03-01Volume 181399141182495Comparative Analysis of the Interaction of Silver Nanoparticles with Hexokinase from Trypanosoma brucei and HumansMlozen MMVan Marwijk JWilhelmi BSWhiteley CMadalitso M Mlozen,1,2 Jacqueline Van Marwijk,1 Brendan Shane Wilhelmi,1 Chris Whiteley1 1Department of Biochemistry and Microbiology, Rhodes University, Makhanda (Grahamstown), South Africa; 2Malawi Adventist University, Malamulo Campus, Department of Biomedical Sciences, Makwasa, MalawiCorrespondence: Madalitso M Mlozen, Malawi Adventist University, Malamulo campus, Department of Biomedical Sciences, P.O.Box 55, Makwasa, Tel +265 884628334, Email mlozenim@mchs.advenist.orgBackground: Regardless of the efforts to ease cases of human African trypanosomiasis (HAT), an increased number of cases get reported annually. This is because of drug resistant Trypanosoma brucei (Tb), the causative agent of the illness. This has renewed the need for creative methods to find new anti-trypanosomal drugs. The blood stream form (BSF) of the parasite depends exclusively on the glycolytic pathway for energy production while it is in the human host. Interruptions in this pathway efficiently kills the parasite. Trypanosoma brucei hexokinase (TbHK) is the first enzyme in glycolysis, and any effectors or inhibitors of TbHK would have potential as anti-trypanosomal agents.Methods: TbHK and human glucokinase (hGCK) were over-expressed with a 6 histidine-tag in E. coli BL21(DE3) cells having the pRARE2 plasmid.Results: TbHK had thermal and pH stability between 30°C and 55°C and 7.5 and 8.5, respectively, while hGCK exhibited thermal and pH stability between 30°C and 40°C and 7.0 and 8.0, respectively. Kinetically, TbHK had a Km of 39.3 μM, Vmax of 0.066 μmol.min− 1.mL− 1, kcat of 2.05 min− 1 and kcat/Km of 0.0526 min− 1.μmol− 1. hGCK exhibited a Km of 4.5 μM, Vmax of 0.032 μnmol.min− 1.mL− 1, kcat of 11.25 min− 1, and kcat/Km of 2.5 min− 1.μmol− 1. Interaction kinetic studies of silver nanoparticles (AgNPs) (0.1 μM) of average size of 6 nm with TbHK and hGCK were conducted. AgNPs selectively inhibited TbHK over hGCK. TbHK showed a non-competitive inhibition with a 50% and 28% decrease in Vmax, and kcat/km, respectively. HsGCK showed a 33% increase in affinity, 9% decrease in Vmax, and a 50% increase in enzyme efficiency.Conclusion: The observed pattern of hGCK and AgNPs falls under the uncompetitive inhibition. The observed highly selective inhibitory effects of AgNPs between TbHK and hGCK may be used in development of new anti-trypanosomal drugs.Keywords: nanoparticles, hexokinase, trypanosomiasis, nanomedicinehttps://www.dovepress.com/comparative-analysis-of-the-interaction-of-silver-nanoparticles-with-h-peer-reviewed-fulltext-article-IJNnanoparticleshexokinasetrypanosomiasisnanomedicine |
| spellingShingle | Mlozen MM Van Marwijk J Wilhelmi BS Whiteley C Comparative Analysis of the Interaction of Silver Nanoparticles with Hexokinase from Trypanosoma brucei and Humans International Journal of Nanomedicine nanoparticles hexokinase trypanosomiasis nanomedicine |
| title | Comparative Analysis of the Interaction of Silver Nanoparticles with Hexokinase from Trypanosoma brucei and Humans |
| title_full | Comparative Analysis of the Interaction of Silver Nanoparticles with Hexokinase from Trypanosoma brucei and Humans |
| title_fullStr | Comparative Analysis of the Interaction of Silver Nanoparticles with Hexokinase from Trypanosoma brucei and Humans |
| title_full_unstemmed | Comparative Analysis of the Interaction of Silver Nanoparticles with Hexokinase from Trypanosoma brucei and Humans |
| title_short | Comparative Analysis of the Interaction of Silver Nanoparticles with Hexokinase from Trypanosoma brucei and Humans |
| title_sort | comparative analysis of the interaction of silver nanoparticles with hexokinase from trypanosoma brucei and humans |
| topic | nanoparticles hexokinase trypanosomiasis nanomedicine |
| url | https://www.dovepress.com/comparative-analysis-of-the-interaction-of-silver-nanoparticles-with-h-peer-reviewed-fulltext-article-IJN |
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