The Experts below are selected from a list of 276 Experts worldwide ranked by ideXlab platform
Michele Vendruscolo - One of the best experts on this subject based on the ideXlab platform.
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Facile Installation of Post-translational Modifications on the Tau Protein via Chemical Mutagenesis.
ACS chemical neuroscience, 2021Co-Authors: Philip R. Lindstedt, Ross J. Taylor, Gonçalo J. L. Bernardes, Michele VendruscoloAbstract:Post-translational modifications of proteins are ubiquitous in living organisms, as they enable an accurate control of the interactions of these macromolecules. For mechanistic studies, it would be highly advantageous to be able to produce in vitro post-translationally modified proteins with site-specificity. Here, we demonstrate one facile way to achieve this goal through the use of post-translational Chemical Mutagenesis. We illustrate this approach by performing site-specific phosphorylation and methylation of tau, a protein that stabilizes microtubules and whose aggregation is closely linked with Alzheimer's disease. We then verify the effects of the post-translational modifications on the ability of tau to control microtubule polymerization, revealing in particular an unexpected role for phosphorylation at S199, which is outside the microtubule-binding region of tau. These results show how the Chemical Mutagenesis approach that we present enables the systematic analysis of site-specific post-translational modifications of a key protein involved in the pathogenesis of Alzheimer's disease.
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Systematic Activity Maturation of a Single-Domain Antibody with Non-canonical Amino Acids through Chemical Mutagenesis.
Cell chemical biology, 2020Co-Authors: Philip R. Lindstedt, Gonçalo J. L. Bernardes, Francesco A. Aprile, Pietro Sormanni, Robertinah Rakoto, Christopher M. Dobson, Michele VendruscoloAbstract:Great advances have been made over the last four decades in therapeutic and diagnostic applications of antibodies. The activity maturation of antibody candidates, however, remains a significant challenge. To address this problem, we present a method that enables the systematic enhancement of the activity of a single-domain antibody through the post-translational installation of non-canonical side chains by Chemical Mutagenesis. We illustrate this approach by performing a structure-activity relationship study beyond the 20 naturally occurring amino acids on a single-domain antibody designed in silico to inhibit the aggregation of the amyloid-β peptide, a process closely linked to Alzheimer's disease. We found that this approach can improve, by five orders of magnitude, the anti-aggregation activity of the starting single-domain antibody, without affecting its stability. These results show that the expansion of the Chemical space available to antibodies through Chemical Mutagenesis can be exploited for the systematic enhancement of the activity of these molecules.
Philip R. Lindstedt - One of the best experts on this subject based on the ideXlab platform.
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Facile Installation of Post-translational Modifications on the Tau Protein via Chemical Mutagenesis.
ACS chemical neuroscience, 2021Co-Authors: Philip R. Lindstedt, Ross J. Taylor, Gonçalo J. L. Bernardes, Michele VendruscoloAbstract:Post-translational modifications of proteins are ubiquitous in living organisms, as they enable an accurate control of the interactions of these macromolecules. For mechanistic studies, it would be highly advantageous to be able to produce in vitro post-translationally modified proteins with site-specificity. Here, we demonstrate one facile way to achieve this goal through the use of post-translational Chemical Mutagenesis. We illustrate this approach by performing site-specific phosphorylation and methylation of tau, a protein that stabilizes microtubules and whose aggregation is closely linked with Alzheimer's disease. We then verify the effects of the post-translational modifications on the ability of tau to control microtubule polymerization, revealing in particular an unexpected role for phosphorylation at S199, which is outside the microtubule-binding region of tau. These results show how the Chemical Mutagenesis approach that we present enables the systematic analysis of site-specific post-translational modifications of a key protein involved in the pathogenesis of Alzheimer's disease.
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Systematic Activity Maturation of a Single-Domain Antibody with Non-canonical Amino Acids through Chemical Mutagenesis.
Cell chemical biology, 2020Co-Authors: Philip R. Lindstedt, Gonçalo J. L. Bernardes, Francesco A. Aprile, Pietro Sormanni, Robertinah Rakoto, Christopher M. Dobson, Michele VendruscoloAbstract:Great advances have been made over the last four decades in therapeutic and diagnostic applications of antibodies. The activity maturation of antibody candidates, however, remains a significant challenge. To address this problem, we present a method that enables the systematic enhancement of the activity of a single-domain antibody through the post-translational installation of non-canonical side chains by Chemical Mutagenesis. We illustrate this approach by performing a structure-activity relationship study beyond the 20 naturally occurring amino acids on a single-domain antibody designed in silico to inhibit the aggregation of the amyloid-β peptide, a process closely linked to Alzheimer's disease. We found that this approach can improve, by five orders of magnitude, the anti-aggregation activity of the starting single-domain antibody, without affecting its stability. These results show that the expansion of the Chemical space available to antibodies through Chemical Mutagenesis can be exploited for the systematic enhancement of the activity of these molecules.
Gonçalo J. L. Bernardes - One of the best experts on this subject based on the ideXlab platform.
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Facile Installation of Post-translational Modifications on the Tau Protein via Chemical Mutagenesis.
ACS chemical neuroscience, 2021Co-Authors: Philip R. Lindstedt, Ross J. Taylor, Gonçalo J. L. Bernardes, Michele VendruscoloAbstract:Post-translational modifications of proteins are ubiquitous in living organisms, as they enable an accurate control of the interactions of these macromolecules. For mechanistic studies, it would be highly advantageous to be able to produce in vitro post-translationally modified proteins with site-specificity. Here, we demonstrate one facile way to achieve this goal through the use of post-translational Chemical Mutagenesis. We illustrate this approach by performing site-specific phosphorylation and methylation of tau, a protein that stabilizes microtubules and whose aggregation is closely linked with Alzheimer's disease. We then verify the effects of the post-translational modifications on the ability of tau to control microtubule polymerization, revealing in particular an unexpected role for phosphorylation at S199, which is outside the microtubule-binding region of tau. These results show how the Chemical Mutagenesis approach that we present enables the systematic analysis of site-specific post-translational modifications of a key protein involved in the pathogenesis of Alzheimer's disease.
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Systematic Activity Maturation of a Single-Domain Antibody with Non-canonical Amino Acids through Chemical Mutagenesis.
Cell chemical biology, 2020Co-Authors: Philip R. Lindstedt, Gonçalo J. L. Bernardes, Francesco A. Aprile, Pietro Sormanni, Robertinah Rakoto, Christopher M. Dobson, Michele VendruscoloAbstract:Great advances have been made over the last four decades in therapeutic and diagnostic applications of antibodies. The activity maturation of antibody candidates, however, remains a significant challenge. To address this problem, we present a method that enables the systematic enhancement of the activity of a single-domain antibody through the post-translational installation of non-canonical side chains by Chemical Mutagenesis. We illustrate this approach by performing a structure-activity relationship study beyond the 20 naturally occurring amino acids on a single-domain antibody designed in silico to inhibit the aggregation of the amyloid-β peptide, a process closely linked to Alzheimer's disease. We found that this approach can improve, by five orders of magnitude, the anti-aggregation activity of the starting single-domain antibody, without affecting its stability. These results show that the expansion of the Chemical space available to antibodies through Chemical Mutagenesis can be exploited for the systematic enhancement of the activity of these molecules.
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Posttranslational Chemical Mutagenesis: To Reveal the Role of Noncatalytic Cysteine Residues in Pathogenic Bacterial Phosphatases.
Biochemistry, 2018Co-Authors: Jb Bertoldo, Hernán Terenzi, Stefan Hüttelmaier, Gonçalo J. L. BernardesAbstract:The field of Chemical site-selective modification of proteins has progressed extensively in recent decades to enable protein functionalization for imaging, drug delivery, and functional studies. In this Perspective, we provide detailed insight into an alternative use of site-selective protein chemistry to probe the role(s) of unpaired Cys residues in the structure and function of disease relevant proteins. Phosphatases are important players in the successful infection of pathogenic bacteria, which represent a significant health burden, particularly in multi-drug-resistant strains. Therefore, a strategy for readily probing the key amino acid role(s) in structure and function may facilitate the targeting and inhibition of these virulence factors. With a dehydroalanine-based posttranslational Chemical Mutagenesis approach, it is possible to reveal hitherto unknown function(s) of noncatalytic Cys residues and confirm their role and interplay in pathogenic bacterial phosphatases. By selectively modifying react...
Cheng-bin Cui - One of the best experts on this subject based on the ideXlab platform.
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seven new and two known lipopeptides as well as five known polyketides the activated production of silent metabolites in a marine derived fungus by Chemical Mutagenesis strategy using diethyl sulphate
Marine Drugs, 2014Co-Authors: Cheng-bin CuiAbstract:AD-2-1 is an antitumor fungal mutant obtained by diethyl sulfate Mutagenesis of a marine-derived Penicillium purpurogenum G59. The G59 strain originally did not produce any metabolites with antitumor activities in MTT assays using K562 cells. Tracing newly produced metabolites under guidance of MTT assay and TLC analysis by direct comparison with control G59 extract, seven new (1–7) and two known (8–9) lipopeptides were isolated together with five known polyketides 10–14 from the extract of mutant AD-2-1. Structures of the seven new compounds including their absolute configurations were determined by spectroscopic and Chemical evidences and named as penicimutalides A–G (1–7). Seven known compounds were identified as fellutamide B (8), fellutamide C (9), 1′-O-methylaverantin (10), averantin (11), averufin (12), nidurufin (13), and sterigmatocystin (14). In the MTT assay, 1–14 inhibited several human cancer cell lines to varying extents. All the bioassays and HPLC-photodiode array detector (PDAD)-UV and HPLC-electron spray ionization (ESI)-MS analyses demonstrated that the production of 1–14 in the mutant AD-2-1 was caused by the activated production of silent metabolites in the original G59 fungal strain. Present results provided additional examples for effectiveness of the Chemical Mutagenesis strategy using diethyl sulphate Mutagenesis to discover new compounds by activating silent metabolites in fungal isolates.
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Three new and eleven known unusual C25 steroids: activated production of silent metabolites in a marine-derived fungus by Chemical Mutagenesis strategy using diethyl sulphate.
Marine drugs, 2014Co-Authors: Ming-wen Xia, Cheng-bin CuiAbstract:Three new (1–3) and 11 known (4–14) C25 steroids with an unusual bicyclo[4.4.1]A/B ring system were isolated by tracing newly produced metabolites in the EtOAc extract of an antitumor mutant AD-1-2 obtained by the diethyl sulphate (DES) Mutagenesis of a marine-derived Penicillium purpurogenum G59. HPLC-PDAD-UV and HPLC-ESI-MS analyses indicated that the G59 strain did not produce these metabolites and the production of 1–14 in the mutant AD-1-2 extract was caused by the activation of silent metabolites in the original G59 strain by DES Mutagenesis. The structures of the new compounds, named antineocyclocitrinols A (1) and B (2) and 23-O-methylantineocyclocitrinol (3), including their absolute configurations were determined by various spectroscopic methods, especially the NMR and Mo2-induced CD analyses. Compounds 1–3 provide the first examples of the C25 bicyclo[4.4.1]A/B ring steroids with the Z-configuration of 20,22-double bond. All of 1–14 weakly inhibited several human cancer cell lines to varying extents. These results provided additional examples for the successful application of the Chemical Mutagenesis strategy using DES to discover new compounds by activating silent metabolites in fungal isolates and supported also the effectiveness and usefulness of this new strategy.
Francesco A. Aprile - One of the best experts on this subject based on the ideXlab platform.
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Systematic Activity Maturation of a Single-Domain Antibody with Non-canonical Amino Acids through Chemical Mutagenesis.
Cell chemical biology, 2020Co-Authors: Philip R. Lindstedt, Gonçalo J. L. Bernardes, Francesco A. Aprile, Pietro Sormanni, Robertinah Rakoto, Christopher M. Dobson, Michele VendruscoloAbstract:Great advances have been made over the last four decades in therapeutic and diagnostic applications of antibodies. The activity maturation of antibody candidates, however, remains a significant challenge. To address this problem, we present a method that enables the systematic enhancement of the activity of a single-domain antibody through the post-translational installation of non-canonical side chains by Chemical Mutagenesis. We illustrate this approach by performing a structure-activity relationship study beyond the 20 naturally occurring amino acids on a single-domain antibody designed in silico to inhibit the aggregation of the amyloid-β peptide, a process closely linked to Alzheimer's disease. We found that this approach can improve, by five orders of magnitude, the anti-aggregation activity of the starting single-domain antibody, without affecting its stability. These results show that the expansion of the Chemical space available to antibodies through Chemical Mutagenesis can be exploited for the systematic enhancement of the activity of these molecules.