Characterization of low molecular weight protein tyrosine phosphatases of Entamoeba histolytica
| Autor: | Francisco Sierra-López, Sonia Cynthia Vanegas-Villa, José Luis Rosales-Encina, Lidia Baylón-Pacheco |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Erythrocytes Phosphatase Protein tyrosine phosphatase Biochemistry Substrate Specificity 03 medical and health sciences chemistry.chemical_compound Entamoeba histolytica Cricetinae Enzyme Stability medicine Animals Humans Amino Acid Sequence Trophozoites Enzyme Inhibitors Tyrosine Chelating Agents chemistry.chemical_classification Mice Inbred BALB C Amoebic liver abscess 030102 biochemistry & molecular biology biology Temperature General Medicine Hydrogen-Ion Concentration medicine.disease biology.organism_classification Recombinant Proteins Amino acid Molecular Weight 030104 developmental biology Enzyme chemistry Phosphoserine Liver Abscess Amebic Female Protein Tyrosine Phosphatases |
| Zdroj: | Biochimie. 180:43-53 |
| ISSN: | 0300-9084 |
| DOI: | 10.1016/j.biochi.2020.10.015 |
| Popis: | Entamoeba histolytica is an intestinal protozoan parasite of humans and is endemic in developing countries. E. histolytica has two low molecular weight protein tyrosine phosphatase (LMW-PTP) genes, EhLMW-PTP1 and EhLMW-PTP2, which are expressed in cultured trophozoites, clinical isolates, and cysts. The amino acid sequences of proteins EhLMW-PTP1 and EhLMW-PTP2 showed only one amino acid difference between them at position A85V, respectively. Both genes are expressed in cultured trophozoites, mainly EhLMW-PTP2, and in trophozoites recovered from amoebic liver abscess, the expression of EhLMW-PTP1 is downregulated. We cloned the two genes and purified the corresponding recombinant (rEhLMW-PTPs) proteins. Antibodies anti-rEhLMW-PTP2 showed that during red blood cells uptake by E. histolytica, the EhLMW-PTPs were found in the phagocytic cups based on analysis of fluorescence signals. On the other hand, rEhLMW-PTPs showed an optimum phosphatase activity at pH 6.0 with p-nitrophenyl phosphate as the substrate. They dephosphorylate phosphotyrosine and 3-O-methylfluorescein phosphate, but not phosphoserine or phosphothreonine, and the enzymatic activity is inhibited by orthovanadate. rEhLMW-PTP1 and rEhLMW-PTP2 exhibited optimum temperatures of activities at 60 °C and 58 °C, respectively, with high thermal stability at 50 °C. Also, the rEhLMW-PTPs showed high specific activities and specific km value with pNPP or OMFP as the substrates at the physiological temperature (37 °C). |
| Databáze: | OpenAIRE |
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