Pseudokinases, Volume 667, the latest release in the Methods in Enzymology serial, highlights new advances in the field with this new volume presenting interesting chapters, including the Production and Purification of the PEAK pseudokinases for structural and functional studies, Structural biology and biophysical characterization of Tribbles pseudokinases, Detecting endogenous TRIB protein expression and its downstream signaling, Analysis of human Tribbles 2 pseudokinase, Expression, purification and examination of ligand-binding to IRAK pseudokinases, Characterization of pseudokinase ILK-mediated actin assembly, Biochemical examination of Titin pseudokinase, Approaches to study pseudokinase conformations, CRISPR editing cell lines for reconstitution studies of pseudokinase function, and much more.
1. Cover image - p. i 2. Title page - p. ii 3. Table of Contents - p. iii 4. Series Page - p. iii 5. Copyright - p. iii 6. Contributors - p. iii 7. Preface - p. xxi 8. Chapter One: Production and purification of the PEAK pseudokinases for structural and functional studies - p. 1 9. Chapter Two: Structural and biophysical characterization of Tribbles homolog 1 - p. 37 10. Chapter Three: Detecting endogenous TRIB2 protein expression by flow cytometry and Western blotting - p. 59 11. Chapter Four: Analysis of human Tribbles 2 (TRIB2) pseudokinase - p. 79 12. Chapter Five: Purification, crystallization and drug screening of the IRAK pseudokinases - p. 101 13. Chapter Six: Characterization of pseudokinase ILK-mediated actin assembly - p. 123 14. Chapter Seven: Production and analysis of titin kinase: Exploiting active/inactive kinase homologs in pseudokinase validation - p. 147 15. Chapter Eight: Co-expression of recombinant RIPK3:MLKL complexes using the baculovirus-insect cell system - p. 183 16. Chapter Nine: CRISPR deletions in cell lines for reconstitution studies of pseudokinase function - p. 229 17. Chapter Ten: Protein engineering strategies to stimulate the functions of bacterial pseudokinases - p. 275 18. Chapter Eleven: Dynamics of protein kinases and pseudokinases by HDX-MS - p. 303 19. Chapter Twelve: Cellular thermal shift assay (CETSA) for determining the drug binding affinity using Ba/F3 clones stably expressing receptor pseudokinases - p. 339 20. Chapter Thirteen: Conformational control and regulation of the pseudokinase KSR via small molecule binding interactions - p. 365 21. Chapter Fourteen: Computational tools and resources for pseudokinase research - p. 403 22. Chapter Fifteen: Methods to assess small molecule allosteric modulators of the STRAD pseudokinase - p. 427 23. Chapter Sixteen: Targeting the HER3 pseudokinase domain with small molecule inhibitors - p. 455 24. Chapter Seventeen: Considerations for studying phosphorylation of the mitotic checkpoint pseudokinase BUBR1 - p. 507 25. Chapter Eighteen: The pseudokinase domain in receptor guanylyl cyclases - p. 541 26. Chapter Nineteen: Methods for discovering catalytic activities for pseudokinases - p. 575 27. Chapter Twenty: Efficient expression, purification, and visualization by cryo-EM of unliganded near full-length HER3 - p. 611 28. Chapter Twenty-One: An effective strategy for ligand-mediated pulldown of the HER2/HER3/NRG1β heterocomplex and cryo-EM structure determination at low sample concentrations - p. 633 29. Chapter Twenty-Two: Enabling pseudokinases as potential drug targets - p. 663 30. Chapter Twenty-Three: Identification and characterization of TYK2 pseudokinase domain stabilizers that allosterically inhibit TYK2 signaling - p. 685 31. Chapter Twenty-Four: A suite of in vitro and in vivo assays for monitoring the activity of the pseudokinase Bud32 - p. 729 32. Chapter Twenty-Five: Dissecting protein function in vivo: Engineering allelic series in mice using CRISPR-Cas9 technology - p. 775
Pseudokinases, Volume 667, the latest release in the Methods in Enzymology serial, highlights new advances in the field with this new volume presenting interesting chapters, including the Production and Purification of the PEAK pseudokinases for structural and functional studies, Structural biology and biophysical characterization of Tribbles pseudokinases, Detecting endogenous TRIB protein expression and its downstream signaling, Analysis of human Tribbles 2 pseudokinase, Expression, purification and examination of ligand-binding to IRAK pseudokinases, Characterization of pseudokinase ILK-mediated actin assembly, Biochemical examination of Titin pseudokinase, Approaches to study pseudokinase conformations, CRISPR editing cell lines for reconstitution studies of pseudokinase function, and much more.
1. Cover image - p. i 2. Title page - p. ii 3. Table of Contents - p. iii 4. Series Page - p. iii 5. Copyright - p. iii 6. Contributors - p. iii 7. Preface - p. xxi 8. Chapter One: Production and purification of the PEAK pseudokinases for structural and functional studies - p. 1 9. Chapter Two: Structural and biophysical characterization of Tribbles homolog 1 - p. 37 10. Chapter Three: Detecting endogenous TRIB2 protein expression by flow cytometry and Western blotting - p. 59 11. Chapter Four: Analysis of human Tribbles 2 (TRIB2) pseudokinase - p. 79 12. Chapter Five: Purification, crystallization and drug screening of the IRAK pseudokinases - p. 101 13. Chapter Six: Characterization of pseudokinase ILK-mediated actin assembly - p. 123 14. Chapter Seven: Production and analysis of titin kinase: Exploiting active/inactive kinase homologs in pseudokinase validation - p. 147 15. Chapter Eight: Co-expression of recombinant RIPK3:MLKL complexes using the baculovirus-insect cell system - p. 183 16. Chapter Nine: CRISPR deletions in cell lines for reconstitution studies of pseudokinase function - p. 229 17. Chapter Ten: Protein engineering strategies to stimulate the functions of bacterial pseudokinases - p. 275 18. Chapter Eleven: Dynamics of protein kinases and pseudokinases by HDX-MS - p. 303 19. Chapter Twelve: Cellular thermal shift assay (CETSA) for determining the drug binding affinity using Ba/F3 clones stably expressing receptor pseudokinases - p. 339 20. Chapter Thirteen: Conformational control and regulation of the pseudokinase KSR via small molecule binding interactions - p. 365 21. Chapter Fourteen: Computational tools and resources for pseudokinase research - p. 403 22. Chapter Fifteen: Methods to assess small molecule allosteric modulators of the STRAD pseudokinase - p. 427 23. Chapter Sixteen: Targeting the HER3 pseudokinase domain with small molecule inhibitors - p. 455 24. Chapter Seventeen: Considerations for studying phosphorylation of the mitotic checkpoint pseudokinase BUBR1 - p. 507 25. Chapter Eighteen: The pseudokinase domain in receptor guanylyl cyclases - p. 541 26. Chapter Nineteen: Methods for discovering catalytic activities for pseudokinases - p. 575 27. Chapter Twenty: Efficient expression, purification, and visualization by cryo-EM of unliganded near full-length HER3 - p. 611 28. Chapter Twenty-One: An effective strategy for ligand-mediated pulldown of the HER2/HER3/NRG1β heterocomplex and cryo-EM structure determination at low sample concentrations - p. 633 29. Chapter Twenty-Two: Enabling pseudokinases as potential drug targets - p. 663 30. Chapter Twenty-Three: Identification and characterization of TYK2 pseudokinase domain stabilizers that allosterically inhibit TYK2 signaling - p. 685 31. Chapter Twenty-Four: A suite of in vitro and in vivo assays for monitoring the activity of the pseudokinase Bud32 - p. 729 32. Chapter Twenty-Five: Dissecting protein function in vivo: Engineering allelic series in mice using CRISPR-Cas9 technology - p. 775
