دورية أكاديمية
Biophysical characterization of two commercially available preparations of the drug containing Escherichia coli L-Asparaginase 2.
| العنوان: | Biophysical characterization of two commercially available preparations of the drug containing Escherichia coli L-Asparaginase 2. |
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| المؤلفون: | de Araújo TS; Pharmaceutical Biotechnology Laboratory - pbiotech, Faculty of Pharmacy, Federal University of Rio de Janeiro, Rio de Janeiro, RJ 21941-902, Brazil; Protein Advanced Biochemistry - PAB, National Center for Structural Biology and Bioimaging - CENABIO, Federal University of Rio de Janeiro, Rio de Janeiro, RJ, 21941-902, Brazil; Institute for Medical Biochemistry Leopoldo DeMeis, Federal University of Rio de Janeiro, Rio de Janeiro, RJ, 21941-902, Brazil., Scapin SMN; National Institute of Metrology, Quality and Technology - INMETRO, Duque de Caxias, RJ 25250-020, Brazil., de Andrade W; Pharmaceutical Biotechnology Laboratory - pbiotech, Faculty of Pharmacy, Federal University of Rio de Janeiro, Rio de Janeiro, RJ 21941-902, Brazil., Fasciotti M; National Institute of Metrology, Quality and Technology - INMETRO, Duque de Caxias, RJ 25250-020, Brazil., de Magalhães MTQ; Macromolecular Biophysics Laboratory (LBM), Department of Biochemistry and Immunology, Federal University of Minas Gerais, Belo Horizonte, Minas Gerais 31270-901, Brazil., Almeida MS; Protein Advanced Biochemistry - PAB, National Center for Structural Biology and Bioimaging - CENABIO, Federal University of Rio de Janeiro, Rio de Janeiro, RJ, 21941-902, Brazil; Institute for Medical Biochemistry Leopoldo DeMeis, Federal University of Rio de Janeiro, Rio de Janeiro, RJ, 21941-902, Brazil. Electronic address: msalmeida@cenabio.ufrj.br., Lima LMTR; Pharmaceutical Biotechnology Laboratory - pbiotech, Faculty of Pharmacy, Federal University of Rio de Janeiro, Rio de Janeiro, RJ 21941-902, Brazil; National Institute of Metrology, Quality and Technology - INMETRO, Duque de Caxias, RJ 25250-020, Brazil. Electronic address: Mauricio@pharma.ufrj.br. |
| المصدر: | Biophysical chemistry [Biophys Chem] 2021 Apr; Vol. 271, pp. 106554. Date of Electronic Publication: 2021 Feb 05. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier Science B.V Country of Publication: Netherlands NLM ID: 0403171 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1873-4200 (Electronic) Linking ISSN: 03014622 NLM ISO Abbreviation: Biophys Chem Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Amsterdam : Elsevier Science B.V Original Publication: Amsterdam, North-Holland Pub. Co. |
| مواضيع طبية MeSH: | Asparaginase/*metabolism , Escherichia coli/*enzymology, Asparaginase/chemistry ; Protein Conformation |
| مستخلص: | The hydrolysis of asparagine and glutamine by L-asparaginase has been used to treat acute lymphoblastic leukemia for over four decades. Each L-asparaginase monomer has a long loop that closes over the active site upon substrate binding, acting as a lid. Here we present a comparative study of two commercially available preparations of the drug containing Escherichia coli L-Asparaginase 2 (EcA2), performed by a comprehensive array of biophysical and biochemical approaches. We report the oligomeric landscape and conformational and dynamic plasticity of E. coli type 2 L-asparaginase present in two different formulations, and its relationship with L-aspartic acid, which is present in Aginasa, but not in Leuginase. The L-Asp present in Aginasa formulation was found to provide to EcA2 a resistance to in vitro proteolysis. EcA2 shows a composition of monomers and oligomers up to tetramers, which is mostly not altered in the presence of L-Asp. Ion-mobility spectrometry-mass spectrometry reveals two conformers for the monomeric EcA2, and that monomeric species has sufficient capacity for selective binding to L-Asp and L-Glu. The N-terminal loop of the EcA2 present in Leuginase, which is part of the active site is disordered, but it gets ordered in the presence of L-Asp, while L-Glu only does so to a limited extent. These data provide new insights on the mechanistic of ligand recognition by EcA2, and the impact of formulation in its conformational diversity landscape. (Copyright © 2021 Elsevier B.V. All rights reserved.) |
| فهرسة مساهمة: | Keywords: Biopharmaceuticals; Cancer therapy; Enzyme mechanism; L-Asparaginase; Leukemia; Ligand-binding protein; Nuclear magnetic resonance (NMR); Protein conformation; Protein crystallization; Protein dynamic |
| المشرفين على المادة: | EC 3.5.1.1 (Asparaginase) |
| تواريخ الأحداث: | Date Created: 20210219 Date Completed: 20211116 Latest Revision: 20211116 |
| رمز التحديث: | 20231215 |
| DOI: | 10.1016/j.bpc.2021.106554 |
| PMID: | 33607531 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1873-4200 |
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| DOI: | 10.1016/j.bpc.2021.106554 |