2014
Maksimoska, Jasna; Segura-Peña, Dario; Cole, Philip A; Marmorstein, Ronen
Structure of the p300 histone acetyltransferase bound to acetyl-coenzyme A and its analogues Journal Article
In: Biochemistry, vol. 53, no. 21, pp. 3415-22, 2014, ISSN: 1520-4995.
Abstract | Links | BibTeX | Tags: Acetyl Coenzyme A, Catalytic Domain, Coenzyme A, Humans, Models, Molecular, p300-CBP Transcription Factors, Protein Binding, Protein Conformation
@article{1299982,
title = {Structure of the p300 histone acetyltransferase bound to acetyl-coenzyme A and its analogues},
author = {Jasna Maksimoska and Dario Segura-Peña and Philip A Cole and Ronen Marmorstein},
doi = {10.1021/bi500380f},
issn = {1520-4995},
year = {2014},
date = {2014-06-01},
journal = {Biochemistry},
volume = {53},
number = {21},
pages = {3415-22},
abstract = {The p300 and CBP transcriptional coactivator paralogs (p300/CBP) regulate a variety of different cellular pathways, in part, by acetylating histones and more than 70 non-histone protein substrates. Mutation, chromosomal translocation, or other aberrant activities of p300/CBP are linked to many different diseases, including cancer. Because of its pleiotropic biological roles and connection to disease, it is important to understand the mechanism of acetyl transfer by p300/CBP, in part so that inhibitors can be more rationally developed. Toward this goal, a structure of p300 bound to a Lys-CoA bisubstrate HAT inhibitor has been previously elucidated, and the enzyme’s catalytic mechanism has been investigated. Nonetheless, many questions underlying p300/CBP structure and mechanism remain. Here, we report a structural characterization of different reaction states in the p300 activity cycle. We present the structures of p300 in complex with an acetyl-CoA substrate, a CoA product, and an acetonyl-CoA inhibitor. A comparison of these structures with the previously reported p300/Lys-CoA complex demonstrates that the conformation of the enzyme active site depends on the interaction of the enzyme with the cofactor, and is not apparently influenced by protein substrate lysine binding. The p300/CoA crystals also contain two poly(ethylene glycol) moieties bound proximal to the cofactor binding site, implicating the path of protein substrate association. The structure of the p300/acetonyl-CoA complex explains the inhibitory and tight binding properties of the acetonyl-CoA toward p300. Together, these studies provide new insights into the molecular basis of acetylation by p300 and have implications for the rational development of new small molecule p300 inhibitors.},
keywords = {Acetyl Coenzyme A, Catalytic Domain, Coenzyme A, Humans, Models, Molecular, p300-CBP Transcription Factors, Protein Binding, Protein Conformation},
pubstate = {published},
tppubtype = {article}
}
The p300 and CBP transcriptional coactivator paralogs (p300/CBP) regulate a variety of different cellular pathways, in part, by acetylating histones and more than 70 non-histone protein substrates. Mutation, chromosomal translocation, or other aberrant activities of p300/CBP are linked to many different diseases, including cancer. Because of its pleiotropic biological roles and connection to disease, it is important to understand the mechanism of acetyl transfer by p300/CBP, in part so that inhibitors can be more rationally developed. Toward this goal, a structure of p300 bound to a Lys-CoA bisubstrate HAT inhibitor has been previously elucidated, and the enzyme’s catalytic mechanism has been investigated. Nonetheless, many questions underlying p300/CBP structure and mechanism remain. Here, we report a structural characterization of different reaction states in the p300 activity cycle. We present the structures of p300 in complex with an acetyl-CoA substrate, a CoA product, and an acetonyl-CoA inhibitor. A comparison of these structures with the previously reported p300/Lys-CoA complex demonstrates that the conformation of the enzyme active site depends on the interaction of the enzyme with the cofactor, and is not apparently influenced by protein substrate lysine binding. The p300/CoA crystals also contain two poly(ethylene glycol) moieties bound proximal to the cofactor binding site, implicating the path of protein substrate association. The structure of the p300/acetonyl-CoA complex explains the inhibitory and tight binding properties of the acetonyl-CoA toward p300. Together, these studies provide new insights into the molecular basis of acetylation by p300 and have implications for the rational development of new small molecule p300 inhibitors.
Chiang, Meng-Jung; Holbert, Marc A; Kalin, Jay H; Ahn, Young-Hoon; Giddens, John; Amin, Mohammed N; Taylor, Martin S; Collins, Samuel L; Chan-Li, Yee; Waickman, Adam; Hsiao, Po-Yuan; Bolduc, David; Leahy, Daniel J; Horton, Maureen R; Wang, Lai-Xi; Powell, Jonathan D; Cole, Philip A
An Fc domain protein-small molecule conjugate as an enhanced immunomodulator Journal Article
In: J Am Chem Soc, vol. 136, no. 9, pp. 3370-3, 2014, ISSN: 1520-5126.
Abstract | Links | BibTeX | Tags: Adenosine, Animals, CD4-Positive T-Lymphocytes, Immunoconjugates, Immunoglobulin Fc Fragments, Immunologic Factors, Mice, Models, Molecular, Phenethylamines, Protein Conformation
@article{1299984,
title = {An Fc domain protein-small molecule conjugate as an enhanced immunomodulator},
author = {Meng-Jung Chiang and Marc A Holbert and Jay H Kalin and Young-Hoon Ahn and John Giddens and Mohammed N Amin and Martin S Taylor and Samuel L Collins and Yee Chan-Li and Adam Waickman and Po-Yuan Hsiao and David Bolduc and Daniel J Leahy and Maureen R Horton and Lai-Xi Wang and Jonathan D Powell and Philip A Cole},
doi = {10.1021/ja5006674},
issn = {1520-5126},
year = {2014},
date = {2014-03-01},
journal = {J Am Chem Soc},
volume = {136},
number = {9},
pages = {3370-3},
abstract = {Proteins as well as small molecules have demonstrated success as therapeutic agents, but their pharmacologic properties sometimes fall short against particular drug targets. Although the adenosine 2a receptor (A(2A)R) has been identified as a promising target for immunotherapy, small molecule A(2A)R agonists have suffered from short pharmacokinetic half-lives and the potential for toxicity by modulating nonimmune pathways. To overcome these limitations, we have tethered the A(2A)R agonist CGS-21680 to the immunoglobulin Fc domain using expressed protein ligation with Sf9 cell secreted protein. The protein small molecule conjugate Fc-CGS retained potent Fc receptor and A(2A)R interactions and showed superior properties as a therapeutic for the treatment of a mouse model of autoimmune pneumonitis. This approach may provide a general strategy for optimizing small molecule therapeutics.},
keywords = {Adenosine, Animals, CD4-Positive T-Lymphocytes, Immunoconjugates, Immunoglobulin Fc Fragments, Immunologic Factors, Mice, Models, Molecular, Phenethylamines, Protein Conformation},
pubstate = {published},
tppubtype = {article}
}
Proteins as well as small molecules have demonstrated success as therapeutic agents, but their pharmacologic properties sometimes fall short against particular drug targets. Although the adenosine 2a receptor (A(2A)R) has been identified as a promising target for immunotherapy, small molecule A(2A)R agonists have suffered from short pharmacokinetic half-lives and the potential for toxicity by modulating nonimmune pathways. To overcome these limitations, we have tethered the A(2A)R agonist CGS-21680 to the immunoglobulin Fc domain using expressed protein ligation with Sf9 cell secreted protein. The protein small molecule conjugate Fc-CGS retained potent Fc receptor and A(2A)R interactions and showed superior properties as a therapeutic for the treatment of a mouse model of autoimmune pneumonitis. This approach may provide a general strategy for optimizing small molecule therapeutics.
Wang, Z.; Cole, P. A.
Catalytic mechanisms and regulation of protein kinases Journal Article
In: Methods Enzymol., vol. 548, pp. 1-21, 2014, ISSN: 1557-7988.
Abstract | Links | BibTeX | Tags: Adenosine Triphosphate, Animals, Biocatalysis, Humans, Models, Molecular, Mutation, Phosphorylation, Post-Translational, Protein Conformation, Protein Kinase Inhibitors, Protein Kinases, Protein Processing, Substrate Specificity
@article{1299974,
title = {Catalytic mechanisms and regulation of protein kinases},
author = {Z. Wang and P. A. Cole},
doi = {10.1016/B978-0-12-397918-6.00001-X},
issn = {1557-7988},
year = {2014},
date = {2014-00-00},
journal = {Methods Enzymol.},
volume = {548},
pages = {1-21},
abstract = {Protein kinases transfer a phosphoryl group from ATP onto target proteins and play a critical role in signal transduction and other cellular processes. Here, we review the kinase kinetic and chemical mechanisms and their application in understanding kinase structure and function. Aberrant kinase activity has been implicated in many human diseases, in particular cancer. We highlight applications of technologies and concepts derived from kinase mechanistic studies that have helped illuminate how kinases are regulated and contribute to pathophysiology.},
keywords = {Adenosine Triphosphate, Animals, Biocatalysis, Humans, Models, Molecular, Mutation, Phosphorylation, Post-Translational, Protein Conformation, Protein Kinase Inhibitors, Protein Kinases, Protein Processing, Substrate Specificity},
pubstate = {published},
tppubtype = {article}
}
Protein kinases transfer a phosphoryl group from ATP onto target proteins and play a critical role in signal transduction and other cellular processes. Here, we review the kinase kinetic and chemical mechanisms and their application in understanding kinase structure and function. Aberrant kinase activity has been implicated in many human diseases, in particular cancer. We highlight applications of technologies and concepts derived from kinase mechanistic studies that have helped illuminate how kinases are regulated and contribute to pathophysiology.
0000
Rodriguez, Gaddiel; Esadze, Alexandre; Weiser, Brian P; Schonhoft, Joseph D; Cole, Philip A; Stivers, James T
Disordered N-Terminal Domain of Human Uracil DNA Glycosylase (hUNG2) Enhances DNA Translocation Journal Article
In: ACS Chem Biol, vol. 12, no. 9, pp. 2260-2263, 0000, ISSN: 1554-8937.
Abstract | Links | BibTeX | Tags: Binding Sites, Biological Transport, DNA, DNA Glycosylases, Humans, Nuclear Localization Signals, Protein Domains
@article{1299939,
title = {Disordered N-Terminal Domain of Human Uracil DNA Glycosylase (hUNG2) Enhances DNA Translocation},
author = {Gaddiel Rodriguez and Alexandre Esadze and Brian P Weiser and Joseph D Schonhoft and Philip A Cole and James T Stivers},
doi = {10.1021/acschembio.7b00521},
issn = {1554-8937},
journal = {ACS Chem Biol},
volume = {12},
number = {9},
pages = {2260-2263},
abstract = {Nuclear human uracil-DNA glycosylase (hUNG2) initiates base excision repair (BER) of genomic uracils generated through misincorporation of dUMP or through deamination of cytosines. Like many human DNA glycosylases, hUNG2 contains an unstructured N-terminal domain that encodes a nuclear localization signal, protein binding motifs, and sites for post-translational modifications. Although the N-terminal domain has minimal effects on DNA binding and uracil excision kinetics, we report that this domain enhances the ability of hUNG2 to translocate on DNA chains as compared to the catalytic domain alone. The enhancement is most pronounced when physiological ion concentrations and macromolecular crowding agents are used. These data suggest that crowded conditions in the human cell nucleus promote the interaction of the N-terminus with duplex DNA during translocation. The increased contact time with the DNA chain likely contributes to the ability of hUNG2 to locate densely spaced uracils that arise during somatic hypermutation and during fluoropyrimidine chemotherapy.},
keywords = {Binding Sites, Biological Transport, DNA, DNA Glycosylases, Humans, Nuclear Localization Signals, Protein Domains},
pubstate = {published},
tppubtype = {article}
}
Nuclear human uracil-DNA glycosylase (hUNG2) initiates base excision repair (BER) of genomic uracils generated through misincorporation of dUMP or through deamination of cytosines. Like many human DNA glycosylases, hUNG2 contains an unstructured N-terminal domain that encodes a nuclear localization signal, protein binding motifs, and sites for post-translational modifications. Although the N-terminal domain has minimal effects on DNA binding and uracil excision kinetics, we report that this domain enhances the ability of hUNG2 to translocate on DNA chains as compared to the catalytic domain alone. The enhancement is most pronounced when physiological ion concentrations and macromolecular crowding agents are used. These data suggest that crowded conditions in the human cell nucleus promote the interaction of the N-terminus with duplex DNA during translocation. The increased contact time with the DNA chain likely contributes to the ability of hUNG2 to locate densely spaced uracils that arise during somatic hypermutation and during fluoropyrimidine chemotherapy.
Cao, Jia; Peng, Jinghua; An, Hongying; He, Qiyi; Boronina, Tatiana; Guo, Shaodong; White, Morris F; Cole, Philip A; He, Ling
Endotoxemia-mediated activation of acetyltransferase P300 impairs insulin signaling in obesity Journal Article
In: Nat Commun, vol. 8, no. 1, pp. 131, 0000, ISSN: 2041-1723.
Abstract | Links | BibTeX | Tags: Animals, Cell Line, E1A-Associated p300 Protein, Endoplasmic Reticulum Stress, Endotoxemia, Gene Expression Profiling, Immunoblotting, Inbred C57BL, Insulin, Insulin Resistance, Lipopolysaccharides, Liver, Male, Membrane Proteins, Mice, Obese, Obesity, Protein-Serine-Threonine Kinases, Receptor, Signal Transduction, Tumor, X-Box Binding Protein 1
@article{1299941,
title = {Endotoxemia-mediated activation of acetyltransferase P300 impairs insulin signaling in obesity},
author = {Jia Cao and Jinghua Peng and Hongying An and Qiyi He and Tatiana Boronina and Shaodong Guo and Morris F White and Philip A Cole and Ling He},
doi = {10.1038/s41467-017-00163-w},
issn = {2041-1723},
journal = {Nat Commun},
volume = {8},
number = {1},
pages = {131},
abstract = {Diabetes and obesity are characterized by insulin resistance and chronic low-grade inflammation. An elevated plasma concentration of lipopolysaccharide (LPS) caused by increased intestinal permeability during diet-induced obesity promotes insulin resistance in mice. Here, we show that LPS induces endoplasmic reticulum (ER) stress and protein levels of P300, an acetyltransferase involved in glucose production. In high-fat diet fed and genetically obese ob/ob mice, P300 translocates from the nucleus into the cytoplasm of hepatocytes. We also demonstrate that LPS activates the transcription factor XBP1 via the ER stress sensor IRE1, resulting in the induction of P300 which, in turn, acetylates IRS1/2, inhibits its association with the insulin receptor, and disrupts insulin signaling. Pharmacological inhibition of P300 acetyltransferase activity by a specific inhibitor improves insulin sensitivity and decreases hyperglycemia in obese mice. We suggest that P300 acetyltransferase activity may be a promising therapeutic target for the treatment of obese patients.Elevated plasma LPS levels have been associated with insulin resistance. Here Cao et al. show that LPS induces ER stress and P300 activity via the XBP1/IRE1 pathway. P300 acetylates IRS1/2 and inhibits its binding with the insulin receptor. The consequent impairment of insulin signaling can be rescued by pharmacological inhibition of P300.},
keywords = {Animals, Cell Line, E1A-Associated p300 Protein, Endoplasmic Reticulum Stress, Endotoxemia, Gene Expression Profiling, Immunoblotting, Inbred C57BL, Insulin, Insulin Resistance, Lipopolysaccharides, Liver, Male, Membrane Proteins, Mice, Obese, Obesity, Protein-Serine-Threonine Kinases, Receptor, Signal Transduction, Tumor, X-Box Binding Protein 1},
pubstate = {published},
tppubtype = {article}
}
Diabetes and obesity are characterized by insulin resistance and chronic low-grade inflammation. An elevated plasma concentration of lipopolysaccharide (LPS) caused by increased intestinal permeability during diet-induced obesity promotes insulin resistance in mice. Here, we show that LPS induces endoplasmic reticulum (ER) stress and protein levels of P300, an acetyltransferase involved in glucose production. In high-fat diet fed and genetically obese ob/ob mice, P300 translocates from the nucleus into the cytoplasm of hepatocytes. We also demonstrate that LPS activates the transcription factor XBP1 via the ER stress sensor IRE1, resulting in the induction of P300 which, in turn, acetylates IRS1/2, inhibits its association with the insulin receptor, and disrupts insulin signaling. Pharmacological inhibition of P300 acetyltransferase activity by a specific inhibitor improves insulin sensitivity and decreases hyperglycemia in obese mice. We suggest that P300 acetyltransferase activity may be a promising therapeutic target for the treatment of obese patients.Elevated plasma LPS levels have been associated with insulin resistance. Here Cao et al. show that LPS induces ER stress and P300 activity via the XBP1/IRE1 pathway. P300 acetylates IRS1/2 and inhibits its binding with the insulin receptor. The consequent impairment of insulin signaling can be rescued by pharmacological inhibition of P300.
Dancy, Beverley M; Cole, Philip A
Correction to Protein Lysine Acetylation by p300/CBP Journal Article
In: Chem Rev, vol. 116, no. 14, pp. 8314, 0000, ISSN: 1520-6890.
@article{1299964,
title = {Correction to Protein Lysine Acetylation by p300/CBP},
author = {Beverley M Dancy and Philip A Cole},
doi = {10.1021/acs.chemrev.6b00351},
issn = {1520-6890},
journal = {Chem Rev},
volume = {116},
number = {14},
pages = {8314},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Chen, Zan; Thomas, Stefani N; Bolduc, David M; Jiang, Xuejun; Zhang, Xiangbin; Wolberger, Cynthia; Cole, Philip A
Enzymatic Analysis of PTEN Ubiquitylation by WWP2 and NEDD4-1 E3 Ligases Journal Article
In: Biochemistry, vol. 55, no. 26, pp. 3658-66, 0000, ISSN: 1520-4995.
Abstract | Links | BibTeX | Tags: Chromatography, Endosomal Sorting Complexes Required for Transport, Humans, Immunoprecipitation, Liquid, Nedd4 Ubiquitin Protein Ligases, Phosphorylation, Post-Translational, Protein Processing, PTEN Phosphohydrolase, Tandem Mass Spectrometry, Ubiquitin, Ubiquitin-Protein Ligases, Ubiquitination, X-Linked Inhibitor of Apoptosis Protein
@article{1299965,
title = {Enzymatic Analysis of PTEN Ubiquitylation by WWP2 and NEDD4-1 E3 Ligases},
author = {Zan Chen and Stefani N Thomas and David M Bolduc and Xuejun Jiang and Xiangbin Zhang and Cynthia Wolberger and Philip A Cole},
doi = {10.1021/acs.biochem.6b00448},
issn = {1520-4995},
journal = {Biochemistry},
volume = {55},
number = {26},
pages = {3658-66},
abstract = {PTEN is a lipid phosphatase that converts phosphatidylinositol 3,4,5-phosphate (PIP3) to phosphatidylinositol 4,5-phosphate (PIP2) and plays a critical role in the regulation of tumor growth. PTEN is subject to regulation by a variety of post-translational modifications, including phosphorylation on a C-terminal cluster of four Ser/Thr residues (380, 382, 383, and 385) and ubiquitylation by various E3 ligases, including NEDD4-1 and WWP2. It has previously been shown that C-terminal phosphorylation of PTEN can increase its cellular half-life. Using in vitro ubiquitin transfer assays, we show that WWP2 is more active than NEDD4-1 in ubiquitylating unphosphorylated PTEN. The mapping of ubiquitylation sites in PTEN by mass spectrometry showed that both NEDD4-1 and WWP2 can target a broad range of Lys residues in PTEN, although NEDD4-1 versus WWP2 showed a stronger preference for ubiquitylating PTEN’s C2 domain. Whereas tetraphosphorylation of PTEN did not significantly affect its ubiquitylation by NEDD4-1, it inhibited PTEN ubiquitylation by WWP2. Single-turnover and pull-down experiments suggested that tetraphosphorylation of PTEN appears to weaken its interaction with WWP2. These studies reveal how the PTEN E3 ligases WWP2 and NEDD4-1 exhibit distinctive properties in Lys selectivity and sensitivity to PTEN phosphorylation. Our findings also provide a molecular mechanism for the connection between PTEN Ser/Thr phosphorylation and PTEN’s cellular stability.},
keywords = {Chromatography, Endosomal Sorting Complexes Required for Transport, Humans, Immunoprecipitation, Liquid, Nedd4 Ubiquitin Protein Ligases, Phosphorylation, Post-Translational, Protein Processing, PTEN Phosphohydrolase, Tandem Mass Spectrometry, Ubiquitin, Ubiquitin-Protein Ligases, Ubiquitination, X-Linked Inhibitor of Apoptosis Protein},
pubstate = {published},
tppubtype = {article}
}
PTEN is a lipid phosphatase that converts phosphatidylinositol 3,4,5-phosphate (PIP3) to phosphatidylinositol 4,5-phosphate (PIP2) and plays a critical role in the regulation of tumor growth. PTEN is subject to regulation by a variety of post-translational modifications, including phosphorylation on a C-terminal cluster of four Ser/Thr residues (380, 382, 383, and 385) and ubiquitylation by various E3 ligases, including NEDD4-1 and WWP2. It has previously been shown that C-terminal phosphorylation of PTEN can increase its cellular half-life. Using in vitro ubiquitin transfer assays, we show that WWP2 is more active than NEDD4-1 in ubiquitylating unphosphorylated PTEN. The mapping of ubiquitylation sites in PTEN by mass spectrometry showed that both NEDD4-1 and WWP2 can target a broad range of Lys residues in PTEN, although NEDD4-1 versus WWP2 showed a stronger preference for ubiquitylating PTEN’s C2 domain. Whereas tetraphosphorylation of PTEN did not significantly affect its ubiquitylation by NEDD4-1, it inhibited PTEN ubiquitylation by WWP2. Single-turnover and pull-down experiments suggested that tetraphosphorylation of PTEN appears to weaken its interaction with WWP2. These studies reveal how the PTEN E3 ligases WWP2 and NEDD4-1 exhibit distinctive properties in Lys selectivity and sensitivity to PTEN phosphorylation. Our findings also provide a molecular mechanism for the connection between PTEN Ser/Thr phosphorylation and PTEN’s cellular stability.
Philip, Philge; Boija, Ann; Vaid, Roshan; Churcher, Allison M; Meyers, David J; Cole, Philip A; Mannervik, Mattias; Stenberg, Per
Erratum to: CBP binding outside of promoters and enhancers in Drosophila melanogaster Journal Article
In: Epigenetics Chromatin, vol. 9, no. 1, pp. 38, 0000, ISSN: 1756-8935.
Abstract | Links | BibTeX | Tags:
@article{1299962,
title = {Erratum to: CBP binding outside of promoters and enhancers in Drosophila melanogaster},
author = {Philge Philip and Ann Boija and Roshan Vaid and Allison M Churcher and David J Meyers and Philip A Cole and Mattias Mannervik and Per Stenberg},
doi = {10.1186/s13072-016-0088-y},
issn = {1756-8935},
journal = {Epigenetics Chromatin},
volume = {9},
number = {1},
pages = {38},
abstract = {[This corrects the article DOI: 10.1186/s13072-015-0042-4.].},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
[This corrects the article DOI: 10.1186/s13072-015-0042-4.].
Hsiao, Po-Yuan; Kalin, Jay H; Sun, Im-Hong; Amin, Mohammed N; Lo, Ying-Chun; Chiang, Meng-Jung; Giddens, John; Sysa-Shah, Polina; Gabrielson, Kathleen; Wang, Lai-Xi; Powell, Jonathan D; Cole, Philip A
An Fc-Small Molecule Conjugate for Targeted Inhibition of the Adenosine 2A Receptor Journal Article
In: Chembiochem, vol. 17, no. 20, pp. 1951-1960, 0000, ISSN: 1439-7633.
Abstract | Links | BibTeX | Tags: Adenosine A2 Receptor Antagonists, Adenosine A2A, Animals, Female, Humans, Immunoglobulin Fab Fragments, Inbred C57BL, Knockout, Male, Mice, Models, Molecular, Molecular Structure, Receptor, Respiratory Tract Infections, Triazines, Triazoles, Vaccinia virus
@article{1299961,
title = {An Fc-Small Molecule Conjugate for Targeted Inhibition of the Adenosine 2A Receptor},
author = {Po-Yuan Hsiao and Jay H Kalin and Im-Hong Sun and Mohammed N Amin and Ying-Chun Lo and Meng-Jung Chiang and John Giddens and Polina Sysa-Shah and Kathleen Gabrielson and Lai-Xi Wang and Jonathan D Powell and Philip A Cole},
doi = {10.1002/cbic.201600337},
issn = {1439-7633},
journal = {Chembiochem},
volume = {17},
number = {20},
pages = {1951-1960},
abstract = {The adenosine A2A receptor (A2A R) is expressed in immune cells, as well as brain and heart tissue, and has been intensively studied as a therapeutic target for multiple disease indications. Inhibitors of the A2A R have the potential for stimulating immune response, which could be valuable for cancer immune surveillance and mounting a response against pathogens. One well-established potent and selective small molecule A2A R antagonist, ZM-241385 (ZM), has a short pharmacokinetic half-life and the potential for systemic toxicity due to A2A R effects in the brain and the heart. In this study, we designed an analogue of ZM and tethered it to the Fc domain of the immunoglobulin IgG3 by using expressed protein ligation. The resulting protein-small molecule conjugate, Fc-ZM, retained high affinity for two Fc receptors: FcγRI and the neonatal Fc receptor, FcRn. In addition, Fc-ZM was a potent A2A R antagonist, as measured by a cell-based cAMP assay. Cell-based assays also revealed that Fc-ZM could stimulate interferon γ production in splenocytes in a fashion that was dependent on the presence of A2A R. We found that Fc-ZM, compared with the small molecule ZM, was a superior A2A R antagonist in mice, consistent with the possibility that Fc attachment can improve pharmacokinetic and/or pharmacodynamic properties of the small molecule.},
keywords = {Adenosine A2 Receptor Antagonists, Adenosine A2A, Animals, Female, Humans, Immunoglobulin Fab Fragments, Inbred C57BL, Knockout, Male, Mice, Models, Molecular, Molecular Structure, Receptor, Respiratory Tract Infections, Triazines, Triazoles, Vaccinia virus},
pubstate = {published},
tppubtype = {article}
}
The adenosine A2A receptor (A2A R) is expressed in immune cells, as well as brain and heart tissue, and has been intensively studied as a therapeutic target for multiple disease indications. Inhibitors of the A2A R have the potential for stimulating immune response, which could be valuable for cancer immune surveillance and mounting a response against pathogens. One well-established potent and selective small molecule A2A R antagonist, ZM-241385 (ZM), has a short pharmacokinetic half-life and the potential for systemic toxicity due to A2A R effects in the brain and the heart. In this study, we designed an analogue of ZM and tethered it to the Fc domain of the immunoglobulin IgG3 by using expressed protein ligation. The resulting protein-small molecule conjugate, Fc-ZM, retained high affinity for two Fc receptors: FcγRI and the neonatal Fc receptor, FcRn. In addition, Fc-ZM was a potent A2A R antagonist, as measured by a cell-based cAMP assay. Cell-based assays also revealed that Fc-ZM could stimulate interferon γ production in splenocytes in a fashion that was dependent on the presence of A2A R. We found that Fc-ZM, compared with the small molecule ZM, was a superior A2A R antagonist in mice, consistent with the possibility that Fc attachment can improve pharmacokinetic and/or pharmacodynamic properties of the small molecule.
Ji, Zhiqin; Clark, Richard F; Bhat, Vikram; Hansen, T Matthew; Lasko, Loren M; Bromberg, Kenneth D; Manaves, Vlasios; Algire, Mikkel; Martin, Ruth; Qiu, Wei; Torrent, Maricel; Jakob, Clarissa G; Liu, Hong; Cole, Philip A; Marmorstein, Ronen; Kesicki, Edward A; Lai, Albert; Michaelides, Michael R
Discovery of spirohydantoins as selective, orally bioavailable inhibitors of p300/CBP histone acetyltransferases Journal Article
In: Bioorg Med Chem Lett, vol. 39, pp. 127854, 0000, ISSN: 1464-3405.
Abstract | Links | BibTeX | Tags: Administration, Biological Availability, CREB-Binding Protein, Dose-Response Relationship, Drug, Drug Discovery, E1A-Associated p300 Protein, Enzyme Inhibitors, Humans, Hydantoins, Molecular Structure, Oral, Spiro Compounds, Structure-Activity Relationship
@article{1624368,
title = {Discovery of spirohydantoins as selective, orally bioavailable inhibitors of p300/CBP histone acetyltransferases},
author = {Zhiqin Ji and Richard F Clark and Vikram Bhat and T Matthew Hansen and Loren M Lasko and Kenneth D Bromberg and Vlasios Manaves and Mikkel Algire and Ruth Martin and Wei Qiu and Maricel Torrent and Clarissa G Jakob and Hong Liu and Philip A Cole and Ronen Marmorstein and Edward A Kesicki and Albert Lai and Michael R Michaelides},
doi = {10.1016/j.bmcl.2021.127854},
issn = {1464-3405},
journal = {Bioorg Med Chem Lett},
volume = {39},
pages = {127854},
abstract = {p300 and CREB-binding protein (CBP) are essential for a multitude of cellular processes. Dysregulation of p300/CBP histone acetyltransferase activity is linked to a broad spectrum of human diseases including cancers. A novel drug-like spirohydantoin (21) has been discovered as a selective orally bioavailable inhibitor of p300/CBP histone acetyltransferase. Lead compound 21 is more potent than the first-in-class lead A-485 in both enzymatic and cellular assays and lacks the off-target inhibition of dopamine and serotonin transporters, that was observed with A-485.},
keywords = {Administration, Biological Availability, CREB-Binding Protein, Dose-Response Relationship, Drug, Drug Discovery, E1A-Associated p300 Protein, Enzyme Inhibitors, Humans, Hydantoins, Molecular Structure, Oral, Spiro Compounds, Structure-Activity Relationship},
pubstate = {published},
tppubtype = {article}
}
p300 and CREB-binding protein (CBP) are essential for a multitude of cellular processes. Dysregulation of p300/CBP histone acetyltransferase activity is linked to a broad spectrum of human diseases including cancers. A novel drug-like spirohydantoin (21) has been discovered as a selective orally bioavailable inhibitor of p300/CBP histone acetyltransferase. Lead compound 21 is more potent than the first-in-class lead A-485 in both enzymatic and cellular assays and lacks the off-target inhibition of dopamine and serotonin transporters, that was observed with A-485.
Aufhauser, David D; Hernandez, Paul; Concors, Seth J; O’Brien, Ciaran; Wang, Zhonglin; Murken, Douglas R; Samanta, Arabinda; Beier, Ulf H; Krumeich, Lauren; Bhatti, Tricia R; Wang, Yanfeng; Ge, Guanghui; Wang, Liqing; Cheraghlou, Shayan; Wagner, Florence F; Holson, Edward B; Kalin, Jay H; Cole, Philip A; Hancock, Wayne W; Levine, Matthew H
HDAC2 targeting stabilizes the CoREST complex in renal tubular cells and protects against renal ischemia/reperfusion injury Journal Article
In: Sci Rep, vol. 11, no. 1, pp. 9018, 0000, ISSN: 2045-2322.
Abstract | Links | BibTeX | Tags: Animals, Co-Repressor Proteins, Endothelins, Enzyme Inhibitors, Female, Gene Deletion, Histone Deacetylase 1, Histone Deacetylase 2, Isoenzymes, Kidney Tubules, Knockout, Male, Mice, Proximal, Reperfusion Injury
@article{1624367,
title = {HDAC2 targeting stabilizes the CoREST complex in renal tubular cells and protects against renal ischemia/reperfusion injury},
author = {David D Aufhauser and Paul Hernandez and Seth J Concors and Ciaran O’Brien and Zhonglin Wang and Douglas R Murken and Arabinda Samanta and Ulf H Beier and Lauren Krumeich and Tricia R Bhatti and Yanfeng Wang and Guanghui Ge and Liqing Wang and Shayan Cheraghlou and Florence F Wagner and Edward B Holson and Jay H Kalin and Philip A Cole and Wayne W Hancock and Matthew H Levine},
doi = {10.1038/s41598-021-88242-3},
issn = {2045-2322},
journal = {Sci Rep},
volume = {11},
number = {1},
pages = {9018},
abstract = {Histone/protein deacetylases (HDAC) 1 and 2 are typically viewed as structurally and functionally similar enzymes present within various co-regulatory complexes. We tested differential effects of these isoforms in renal ischemia reperfusion injury (IRI) using inducible knockout mice and found no significant change in ischemic tolerance with HDAC1 deletion, but mitigation of ischemic injury with HDAC2 deletion. Restriction of HDAC2 deletion to the kidney via transplantation or PAX8-controlled proximal renal tubule-specific Cre resulted in renal IRI protection. Pharmacologic inhibition of HDAC2 increased histone acetylation in the kidney but did not extend renal protection. Protein analysis demonstrated increased HDAC1-associated CoREST protein in HDAC2-/- versus WT cells, suggesting that in the absence of HDAC2, increased CoREST complex occupancy of HDAC1 can stabilize this complex. In vivo administration of a CoREST inhibitor exacerbated renal injury in WT mice and eliminated the benefit of HDAC2 deletion. Gene expression analysis of endothelin showed decreased endothelin levels in HDAC2 deletion. These data demonstrate that contrasting effects of HDAC1 and 2 on CoREST complex stability within renal tubules can affect outcomes of renal IRI and implicate endothelin as a potential downstream mediator.},
keywords = {Animals, Co-Repressor Proteins, Endothelins, Enzyme Inhibitors, Female, Gene Deletion, Histone Deacetylase 1, Histone Deacetylase 2, Isoenzymes, Kidney Tubules, Knockout, Male, Mice, Proximal, Reperfusion Injury},
pubstate = {published},
tppubtype = {article}
}
Histone/protein deacetylases (HDAC) 1 and 2 are typically viewed as structurally and functionally similar enzymes present within various co-regulatory complexes. We tested differential effects of these isoforms in renal ischemia reperfusion injury (IRI) using inducible knockout mice and found no significant change in ischemic tolerance with HDAC1 deletion, but mitigation of ischemic injury with HDAC2 deletion. Restriction of HDAC2 deletion to the kidney via transplantation or PAX8-controlled proximal renal tubule-specific Cre resulted in renal IRI protection. Pharmacologic inhibition of HDAC2 increased histone acetylation in the kidney but did not extend renal protection. Protein analysis demonstrated increased HDAC1-associated CoREST protein in HDAC2-/- versus WT cells, suggesting that in the absence of HDAC2, increased CoREST complex occupancy of HDAC1 can stabilize this complex. In vivo administration of a CoREST inhibitor exacerbated renal injury in WT mice and eliminated the benefit of HDAC2 deletion. Gene expression analysis of endothelin showed decreased endothelin levels in HDAC2 deletion. These data demonstrate that contrasting effects of HDAC1 and 2 on CoREST complex stability within renal tubules can affect outcomes of renal IRI and implicate endothelin as a potential downstream mediator.