Development of a Protein Scaffold for Arginine Sensing Generated through the Dissection of the Arginine-Binding Protein from <i>Thermotoga maritima</i>
Arginine is one of the most important nutrients of living organisms as it plays a major role in important biological pathways. However, the accumulation of arginine as consequence of metabolic defects causes hyperargininemia, an autosomal recessive disorder. Therefore, the efficient detection of the...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-10-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/21/20/7503 |
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| author | Giovanni Smaldone Alessia Ruggiero Nicole Balasco Luigi Vitagliano |
| author_facet | Giovanni Smaldone Alessia Ruggiero Nicole Balasco Luigi Vitagliano |
| author_sort | Giovanni Smaldone |
| collection | DOAJ |
| description | Arginine is one of the most important nutrients of living organisms as it plays a major role in important biological pathways. However, the accumulation of arginine as consequence of metabolic defects causes hyperargininemia, an autosomal recessive disorder. Therefore, the efficient detection of the arginine is a field of relevant biomedical/biotechnological interest. Here, we developed protein variants suitable for arginine sensing by mutating and dissecting the multimeric and multidomain structure of <i>Thermotoga maritima</i> arginine-binding protein (TmArgBP). Indeed, previous studies have shown that TmArgBP domain-swapped structure can be manipulated to generate simplified monomeric and single domain scaffolds. On both these stable scaffolds, to measure tryptophan fluorescence variations associated with the arginine binding, a Phe residue of the ligand binding pocket was mutated to Trp. Upon arginine binding, both mutants displayed a clear variation of the Trp fluorescence. Notably, the single domain scaffold variant exhibited a good affinity (~3 µM) for the ligand. Moreover, the arginine binding to this variant could be easily reverted under very mild conditions. Atomic-level data on the recognition process between the scaffold and the arginine were obtained through the determination of the crystal structure of the adduct. Collectively, present data indicate that TmArgBP scaffolds represent promising candidates for developing arginine biosensors. |
| first_indexed | 2024-03-10T15:41:50Z |
| format | Article |
| id | doaj.art-d97703977f9540339b96fdb3fa988ca8 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T15:41:50Z |
| publishDate | 2020-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-d97703977f9540339b96fdb3fa988ca82023-11-20T16:46:05ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-10-012120750310.3390/ijms21207503Development of a Protein Scaffold for Arginine Sensing Generated through the Dissection of the Arginine-Binding Protein from <i>Thermotoga maritima</i>Giovanni Smaldone0Alessia Ruggiero1Nicole Balasco2Luigi Vitagliano3IRCCS SDN, Via Emanuele Gianturco, 113 80143 Naples, ItalyInstitute of Biostructures and Bioimaging, CNR, Via Mezzocannone 16. I-80134 Naples, ItalyInstitute of Biostructures and Bioimaging, CNR, Via Mezzocannone 16. I-80134 Naples, ItalyInstitute of Biostructures and Bioimaging, CNR, Via Mezzocannone 16. I-80134 Naples, ItalyArginine is one of the most important nutrients of living organisms as it plays a major role in important biological pathways. However, the accumulation of arginine as consequence of metabolic defects causes hyperargininemia, an autosomal recessive disorder. Therefore, the efficient detection of the arginine is a field of relevant biomedical/biotechnological interest. Here, we developed protein variants suitable for arginine sensing by mutating and dissecting the multimeric and multidomain structure of <i>Thermotoga maritima</i> arginine-binding protein (TmArgBP). Indeed, previous studies have shown that TmArgBP domain-swapped structure can be manipulated to generate simplified monomeric and single domain scaffolds. On both these stable scaffolds, to measure tryptophan fluorescence variations associated with the arginine binding, a Phe residue of the ligand binding pocket was mutated to Trp. Upon arginine binding, both mutants displayed a clear variation of the Trp fluorescence. Notably, the single domain scaffold variant exhibited a good affinity (~3 µM) for the ligand. Moreover, the arginine binding to this variant could be easily reverted under very mild conditions. Atomic-level data on the recognition process between the scaffold and the arginine were obtained through the determination of the crystal structure of the adduct. Collectively, present data indicate that TmArgBP scaffolds represent promising candidates for developing arginine biosensors.https://www.mdpi.com/1422-0067/21/20/7503protein scaffoldsargininemia diagnosisbiosensorscrystal structureprotein dissectionprotein stability |
| spellingShingle | Giovanni Smaldone Alessia Ruggiero Nicole Balasco Luigi Vitagliano Development of a Protein Scaffold for Arginine Sensing Generated through the Dissection of the Arginine-Binding Protein from <i>Thermotoga maritima</i> International Journal of Molecular Sciences protein scaffolds argininemia diagnosis biosensors crystal structure protein dissection protein stability |
| title | Development of a Protein Scaffold for Arginine Sensing Generated through the Dissection of the Arginine-Binding Protein from <i>Thermotoga maritima</i> |
| title_full | Development of a Protein Scaffold for Arginine Sensing Generated through the Dissection of the Arginine-Binding Protein from <i>Thermotoga maritima</i> |
| title_fullStr | Development of a Protein Scaffold for Arginine Sensing Generated through the Dissection of the Arginine-Binding Protein from <i>Thermotoga maritima</i> |
| title_full_unstemmed | Development of a Protein Scaffold for Arginine Sensing Generated through the Dissection of the Arginine-Binding Protein from <i>Thermotoga maritima</i> |
| title_short | Development of a Protein Scaffold for Arginine Sensing Generated through the Dissection of the Arginine-Binding Protein from <i>Thermotoga maritima</i> |
| title_sort | development of a protein scaffold for arginine sensing generated through the dissection of the arginine binding protein from i thermotoga maritima i |
| topic | protein scaffolds argininemia diagnosis biosensors crystal structure protein dissection protein stability |
| url | https://www.mdpi.com/1422-0067/21/20/7503 |
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