Structure–Function Relationship Study of a Secretory Amoebic Phosphatase: A Computational-Experimental Approach
Phosphatases are hydrolytic enzymes that cleave the phosphoester bond of numerous substrates containing phosphorylated residues. The typical classification divides them into acid or alkaline depending on the pH at which they have optimal activity. The histidine phosphatase (HP) superfamily is a larg...
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MDPI AG
2021-02-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/4/2164 |
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| author | Celina Terán-Ramírez Rosa E. Mares-Alejandre Ana L. Estrada-González Patricia L. A. Muñoz-Muñoz Marco A. Ramos-Ibarra |
| author_facet | Celina Terán-Ramírez Rosa E. Mares-Alejandre Ana L. Estrada-González Patricia L. A. Muñoz-Muñoz Marco A. Ramos-Ibarra |
| author_sort | Celina Terán-Ramírez |
| collection | DOAJ |
| description | Phosphatases are hydrolytic enzymes that cleave the phosphoester bond of numerous substrates containing phosphorylated residues. The typical classification divides them into acid or alkaline depending on the pH at which they have optimal activity. The histidine phosphatase (HP) superfamily is a large group of functionally diverse enzymes characterized by having an active-site His residue that becomes phosphorylated during catalysis. HP enzymes are relevant biomolecules due to their current and potential application in medicine and biotechnology. <i>Entamoeba histolytica</i>, the causative agent of human amoebiasis, contains a gene (<i>EHI_146950</i>) that encodes a putative secretory acid phosphatase (<i>Eh</i>HAPp49), exhibiting sequence similarity to histidine acid phosphatase (HAP)/phytase enzymes, i.e., branch-2 of HP superfamily. To assess whether it has the potential as a biocatalyst in removing phosphate groups from natural substrates, we studied the <i>Eh</i>HAPp49 structural and functional features using a computational-experimental approach. Although the combined outcome of computational analyses confirmed its structural similarity with HP branch-2 proteins, the experimental results showed that the recombinant enzyme (r<i>Eh</i>HAPp49) has negligible HAP/phytase activity. Nonetheless, results from supplementary activity evaluations revealed that r<i>Eh</i>HAPp49 exhibits Mg<sup>2+</sup>-dependent alkaline pyrophosphatase activity. To our knowledge, this study represents the first computational-experimental characterization of <i>Eh</i>HAPp49, which offers further insights into the structure–function relationship and the basis for future research. |
| first_indexed | 2024-03-09T00:38:16Z |
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| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T00:38:16Z |
| publishDate | 2021-02-01 |
| publisher | MDPI AG |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-33f8b8684c9241e988ff291e77b83c1c2023-12-11T17:58:09ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-02-01224216410.3390/ijms22042164Structure–Function Relationship Study of a Secretory Amoebic Phosphatase: A Computational-Experimental ApproachCelina Terán-Ramírez0Rosa E. Mares-Alejandre1Ana L. Estrada-González2Patricia L. A. Muñoz-Muñoz3Marco A. Ramos-Ibarra4Biotechnology and Biosciences Research Group, Faculty of Chemical Sciences and Engineering, Autonomous University of Baja California, Tijuana 22390, MexicoBiotechnology and Biosciences Research Group, Faculty of Chemical Sciences and Engineering, Autonomous University of Baja California, Tijuana 22390, MexicoBiotechnology and Biosciences Research Group, Faculty of Chemical Sciences and Engineering, Autonomous University of Baja California, Tijuana 22390, MexicoBiotechnology and Biosciences Research Group, Faculty of Chemical Sciences and Engineering, Autonomous University of Baja California, Tijuana 22390, MexicoBiotechnology and Biosciences Research Group, Faculty of Chemical Sciences and Engineering, Autonomous University of Baja California, Tijuana 22390, MexicoPhosphatases are hydrolytic enzymes that cleave the phosphoester bond of numerous substrates containing phosphorylated residues. The typical classification divides them into acid or alkaline depending on the pH at which they have optimal activity. The histidine phosphatase (HP) superfamily is a large group of functionally diverse enzymes characterized by having an active-site His residue that becomes phosphorylated during catalysis. HP enzymes are relevant biomolecules due to their current and potential application in medicine and biotechnology. <i>Entamoeba histolytica</i>, the causative agent of human amoebiasis, contains a gene (<i>EHI_146950</i>) that encodes a putative secretory acid phosphatase (<i>Eh</i>HAPp49), exhibiting sequence similarity to histidine acid phosphatase (HAP)/phytase enzymes, i.e., branch-2 of HP superfamily. To assess whether it has the potential as a biocatalyst in removing phosphate groups from natural substrates, we studied the <i>Eh</i>HAPp49 structural and functional features using a computational-experimental approach. Although the combined outcome of computational analyses confirmed its structural similarity with HP branch-2 proteins, the experimental results showed that the recombinant enzyme (r<i>Eh</i>HAPp49) has negligible HAP/phytase activity. Nonetheless, results from supplementary activity evaluations revealed that r<i>Eh</i>HAPp49 exhibits Mg<sup>2+</sup>-dependent alkaline pyrophosphatase activity. To our knowledge, this study represents the first computational-experimental characterization of <i>Eh</i>HAPp49, which offers further insights into the structure–function relationship and the basis for future research.https://www.mdpi.com/1422-0067/22/4/2164HAP/phytase-like phosphataserecombinant protein productionstructure–function characterizationhomology-based modeling<i>Entamoeba histolytica</i> |
| spellingShingle | Celina Terán-Ramírez Rosa E. Mares-Alejandre Ana L. Estrada-González Patricia L. A. Muñoz-Muñoz Marco A. Ramos-Ibarra Structure–Function Relationship Study of a Secretory Amoebic Phosphatase: A Computational-Experimental Approach International Journal of Molecular Sciences HAP/phytase-like phosphatase recombinant protein production structure–function characterization homology-based modeling <i>Entamoeba histolytica</i> |
| title | Structure–Function Relationship Study of a Secretory Amoebic Phosphatase: A Computational-Experimental Approach |
| title_full | Structure–Function Relationship Study of a Secretory Amoebic Phosphatase: A Computational-Experimental Approach |
| title_fullStr | Structure–Function Relationship Study of a Secretory Amoebic Phosphatase: A Computational-Experimental Approach |
| title_full_unstemmed | Structure–Function Relationship Study of a Secretory Amoebic Phosphatase: A Computational-Experimental Approach |
| title_short | Structure–Function Relationship Study of a Secretory Amoebic Phosphatase: A Computational-Experimental Approach |
| title_sort | structure function relationship study of a secretory amoebic phosphatase a computational experimental approach |
| topic | HAP/phytase-like phosphatase recombinant protein production structure–function characterization homology-based modeling <i>Entamoeba histolytica</i> |
| url | https://www.mdpi.com/1422-0067/22/4/2164 |
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