Engineering methionine γ-lyase from Citrobacter freundii for anticancer activity
| العنوان: | Engineering methionine γ-lyase from Citrobacter freundii for anticancer activity |
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| المؤلفون: | Barbara Giabbai, Loredano Pollegioni, Serena Galati, Edi Gabellieri, Vitalia V. Kulikova, Andrea Mozzarelli, Alexey D. Nikulin, Chiara Cocconcelli, Patrizia Cioni, S.V. Revtovich, Nicola Demitri, Serena Faggiano, Samanta Raboni, Annamaria Buschini, Tatyana V. Demidkina, Paola Storici, Elena Rosini, Elena A. Morozova |
| المصدر: | Biochimica et biophysica acta. Proteins and proteomics 1866 (2018): 1260–1270. doi:10.1016/j.bbapap.2018.09.011 info:cnr-pdr/source/autori:Raboni S, Revtovich S, Demitri N, Giabbai B, Storici P, Cocconcelli C, Faggiano S, Rosini E, Pollegioni L, Galati S, Buschini A, Morozova E, Kulikova V, Nikulin A, Gabellieri E, Cioni P, Demidkina T, Mozzarelli A/titolo:Engineering methionine ?-lyase from Citrobacter freundii for anticancer activity/doi:10.1016%2Fj.bbapap.2018.09.011/rivista:Biochimica et biophysica acta. Proteins and proteomics/anno:2018/pagina_da:1260/pagina_a:1270/intervallo_pagine:1260–1270/volume:1866 |
| سنة النشر: | 2018 |
| مصطلحات موضوعية: | 0301 basic medicine, Cancer therapy, Pyridoxal 5′-phosphate dependent enzyme, Biophysics, Antineoplastic Agents, Crystallography, X-Ray, Protein Engineering, Biochemistry, Analytical Chemistry, 03 medical and health sciences, chemistry.chemical_compound, Cytotoxicity activity, Methionine depletion, Protein engineering, Site-saturation mutagenesis, Molecular Biology, Bacterial Proteins, Catalytic Domain, Cell Proliferation, chemistry.chemical_classification, Methionine, biology, Mutagenesis, Wild type, Active site, Lyase, biology.organism_classification, Recombinant Proteins, Citrobacter freundii, Amino acid, Protein Structure, Tertiary, Pyridoxal 5?-phosphate dependent enzyme, Carbon-Sulfur Lyases, Kinetics, 030104 developmental biology, Enzyme, chemistry, biology.protein, Mutagenesis, Site-Directed |
| الوصف: | Methionine deprivation of cancer cells, which are deficient in methionine biosynthesis, has been envisioned as a therapeutic strategy to reduce cancer cell viability. Methionine ?-lyase (MGL), an enzyme that degrades methionine, has been exploited to selectively remove the amino acid from cancer cell environment. In order to increase MGL catalytic activity, we performed sequence and structure conservation analysis of MGLs from various microorganisms. Whereas most of the residues in the active site and at the dimer interface were found to be conserved, residues located in the C-terminal flexible loop, forming a wall of the active site entry channel, were found to be variable. Therefore, we carried out site-saturation mutagenesis at four independent positions of the C-terminal flexible loop, P357, V358, P360 and A366 of MGL from Citrobacter freundii, generating libraries that were screened for activity. Among the active variants, V358Y exhibits a 1.9-fold increase in the catalytic rate and a 3-fold increase in KM, resulting in a catalytic efficiency similar to wild type MGL. V358Y cytotoxic activity was assessed towards a panel of cancer and nonmalignant cell lines and found to exhibit IC50 lower than the wild type. The comparison of the 3D-structure of V358Y MGL with other MGL available structures indicates that the C-terminal loop is either in an open or closed conformation that does not depend on the amino acid at position 358. Nevertheless, mutations at this position allosterically affects catalysis. |
| تدمد: | 1878-1454 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::10b32f97c491f662fb024aa42c23c490Test https://pubmed.ncbi.nlm.nih.gov/30268810Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....10b32f97c491f662fb024aa42c23c490 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 18781454 |
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