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US 12,510,549 B2Granted
Reagents and methods for detecting protein lysine 2-hydroxyisobutyrylation
AssigneePTM Biolab Inc.
InventorsYingming Zhao, Lunzhi Dai
Life of this patent
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Abstract
The invention provides an isolated peptide comprising a lysine 2-hydroxyisobutyrylation site, a lysine 2-hydroxyisobutyrylation specific affinity reagent that specifically binds to the peptide, and a method for detecting protein lysine 2-hydroxyisobutyrylation in a sample using the reagent.
Claims4
A method comprising:
- (a) contacting a histone protein or a fragment thereof with an isolated affinity reagent, wherein the histone protein or a fragment thereof comprises an amino acid sequence selected from the group consisting of DAVTYTEHAK2ohibuR (SEQ ID NO:
- 29), ASGPPVSELITK2ohibuAVAAS (SEQ ID NO:
- 30), AVAASK2ohibuER (SEQ ID NO:
- 31), SGVSLAALK2ohibuK (SEQ ID NO:
- 32), K2ohibuALAAAGYDVEK (SEQ ID NO:
- 33), LGLK2ohibuSLVSK (SEQ ID NO:
- 34), propLGLK2ohibuSLVSKprop (SEQ ID NO:
- 34), SLVSK2ohibuGTLVQTK (SEQ ID NO:
- 35), GTLVQTK2ohibuGTGASGSFK (SEQ ID NO:
- 36), NDEELNK2ohibuLLGK (SEQ ID NO:
- 37), PEPSK2ohibuSAPAPK (SEQ ID NO:
- 38), VLK2ohibuQVHPDTGISSK (SEQ ID NO:
- 39), LAHYNK2ohibuR (SEQ ID NO:
- 40), LLLPGELAK2ohibuHAVSEGTK (SEQ ID NO:
- 41), HAVSEGTK2ohibuAVTK (SEQ ID NO:
- 42), AVTK2ohibuYTSSK (SEQ ID NO:
- 43), QLATK2ohibuAAR (SEQ ID NO:
- 44), YQK2ohibuSTELLIR (SEQ ID NO:
- 45), EIAQDFK2ohibuTDLR (SEQ ID NO:
- 46), VTIMPK2ohibuDIQLAR (SEQ ID NO:
- 47), DNIQGITK2ohibuPAIR (SEQ ID NO:
- 48), K2ohibu TVTAMDVVYALK (SEQ ID NO:
- 49), TVTAMDVVYALK2ohibuR (SEQ ID NO:
- 50), KpropK2ohibuAAKpropKpropPAGVR (SEQ ID NO:
- 51), KpropKpropAAK2ohibuKpropPAGVR (SEQ ID NO:
- 51), KpropAAKpropK2ohibuPAGVR (SEQ ID NO:
- 52), K2ohibuASGPPVSELITKpropAVAASK2ohibu (SEQ ID NO:
- 53), KpropASGPPVSELITKpropAVAASK2ohibuER (SEQ ID NO:
- 54), ALAAAGYDVEK2ohibuNNSR (SEQ ID NO:
- 55), IK2ohibuLGLK (SEQ ID NO:
- 56), propGILVQTK2ohibuGTGASGSFKprop (SEQ ID NO:
- 57), propK2ohibuAASGEAKpropPQAKprop (SEQ ID NO:
- 58), propAASGEAK2ohibuPQAKprop (SEQ ID NO:
- 59), propAASGEAKpropPQAK2ohibuKprop (SEQ ID NO:
- 60), propAK2ohibuKpropPAGAAKprop (SEQ ID NO:
- 61), propAKpropKpropPAGAAK2ohibuKpropPKprop (SEQ ID NO:
- 62), propKpropATGAATPK2ohibuKprop (SEQ ID NO:
- 63), propKpropAK2ohibuKpropPAAAAVTKprop (SEQ ID NO:
- 64), propKpropPAAAAVTK2ohibuKprop (SEQ ID NO:
- 65), KpropVAAKpropKpropK2ohibu (SEQ ID NO:
- 66), GK2ohibuQGGK2ohibuAR (SEQ ID NO:
- 67), K2ohibuGNYSER (SEQ ID NO:
- 68), DNK2ohibuK2ohibuTR (SEQ ID NO:
- 69), NDEELNK2ohibuLLGR (SEQ ID NO:
- 70), VTIAQGGVLPNIQAVLLPK2ohibuK (SEQ ID NO:
- 71), propPEPAK2ohibuSAPAPKprop (SEQ ID NO:
- 72), PEPAK2ohibuSAPAPK (SEQ ID NO:
- 72), propPEPAKpropSAPAPKpropK2ohibuGSK (SEQ ID NO:
- 73), KpropAISK2ohibuAQKprop (SEQ ID NO:
- 74), AVTKpropAQK2ohibuKpropDGKpropKpropR (SEQ ID NO:
- 75), AVTKpropAQKpropK2ohibuDGKpropKpropR (SEQ ID NO:
- 75), K2ohibuESYSVYVYK (SEQ ID NO:
- 76), KESYSVYVYK2ohibuVLK (SEQ ID NO:
- 77), QVHPDTGISSK2ohibuAMGIMNSFVNDIFER (SEQ ID NO:
- 78), TK2ohibuQTAR (SEQ ID NO:
- 79), K2ohibuSTGGKacAPR (SEQ ID NO:
- 80), KpropSTGGK2ohibuAPR (SEQ ID NO:
- 80), K2ohibuQLATKacAAR (SEQ ID NO:
- 81), KQLATK2ohibuAAR (SEQ ID NO:
- 81), KacQLATK2ohibuAAR (SEQ ID NO:
- 81), K2ohibuSAPATGGVKpropKpropPHR (SEQ ID NO:
- 82), KpropSAPATGGVK2ohibuKpropPHR (SEQ ID NO:
- 82), K2ohibu LPFQR (SEQ ID NO:
- 83), K2ohibuGGKacGLGKacGGAKacR (SEQ ID NO:
- 84), GKacGGK2ohibuGLGKacGGAKacR (SEQ ID NO:
- 85), GLGK2ohibuGGAKacR (SEQ ID NO:
- 86), GGKpropGLGKpropGGAK2ohibuR (SEQ ID NO:
- 87), GGK2ohibuGLGKacGGAKacR (SEQ ID NO:
- 87), GGKacGLGK2ohibuGGAKacR (SEQ ID NO:
- 87), RGGVK2ohibuR (SEQ ID NO:
- 88), GVLK2ohibuVFLENVIR (SEQ ID NO:
- 89), KTVTAMDVVYALK2ohibuR (SEQ ID NO:
- 90), AGSAAK2ohibuASQSR (SEQ ID NO:
- 91), K2ohibuETYSSYIYK (SEQ ID NO:
- 92), ETYSSYIYK2ohibuVLK (SEQ ID NO:
- 93), IATEASK2ohibuLAAYNIK (SEQ ID NO:
- 94), LILPGELAK2ohibuHAVSEGTR (SEQ ID NO:
- 95), FQK2ohibuSTELLIR (SEQ ID NO:
- 96), DSVTYTEHAK2ohibuR (SEQ ID NO:
- 97), K2ohibuTVTSLDVVYALK (SEQ ID NO:
- 98), TVTSLDVVYALK2ohibuR (SEQ ID NO:
- 99), GMGK2ohibuVGAK (SEQ ID NO:
- 100), K2ohibuSAPATGGIK (SEQ ID NO:
- 101), YQK2ohibuSTDLLIR (SEQ ID NO:
- 102), VTIMTK2ohibuDMQLAR (SEQ ID NO:
- 103), K2ohibuAPAAAAEK (SEQ ID NO:
- 104), KacAPAAAAEK2ohibuK (SEQ ID NO:
- 105), VLK2ohibuQVHPDVGISK (SEQ ID NO:
- 106), IALESSK2ohibuLVR (SEQ ID NO:
- 107), HAISEGTK2ohibuAVTK (SEQ ID NO:
- 108), AVTK2ohibuFSSSTN (SEQ ID NO:
- 109), TASSK2ohibuQVSR (SEQ ID NO:
- 110), AGGKacAGK2ohibuDSGK (SEQ ID NO:
- 111), AGGK2ohibuAGKacDSGK (SEQ ID NO:
- 111) and SAPAPK2ohibuKacGSK (SEQ ID NO:
- 112), wherein K2ohibu is a 2-hydroxyisobutyrylated lysine, Kprop is a propanoylated lysine, and Kac is an acetylated lysine, wherein the isolated affinity reagent is a pan anti-2-hydroxyisobutyrylated lysine (K2ohibu) antibody, whereby the isolated affinity reagent and the histone protein or a fragment thereof form a binding complex;
- and
- (b) detecting the binding complex.
95 more elements
A method comprising:
- (a) contacting a histone protein or a fragment thereof with an isolated affinity reagent, wherein the histone protein or a fragment thereof comprises an amino acid sequence selected from the group consisting of DAVTYTEHAK2ohibuR (SEQ ID NO:
- 29), ASGPPVSELITK2ohibuAVAAS (SEQ ID NO:
- 30), AVAASK2ohibuER (SEQ ID NO:
- 31), SGVSLAALK2ohibuK (SEQ ID NO:
- 32), K2ohibuALAAAGYDVEK (SEQ ID NO:
- 33), LGLK2ohibuSLVSK (SEQ ID NO:
- 34), propLGLK2ohibuSLVSKprop (SEQ ID NO:
- 34), SLVSK2ohibuGTLVQTK (SEQ ID NO:
- 35), GTLVQTK2ohibuGTGASGSFK (SEQ ID NO:
- 36), NDEELNK2ohibuLLGK (SEQ ID NO:
- 37), PEPSK2ohibuSAPAPK (SEQ ID NO:
- 38), VLK2ohibuQVHPDTGISSK (SEQ ID NO:
- 39), LAHYNK2ohibuR (SEQ ID NO:
- 40), LLLPGELAK2ohibuHAVSEGTK (SEQ ID NO:
- 41), HAVSEGTK2ohibuAVTK (SEQ ID NO:
- 42), AVTK2ohibuYTSSK (SEQ ID NO:
- 43), QLATK2ohibuAAR (SEQ ID NO:
- 44), YQK2ohibuSTELLIR (SEQ ID NO:
- 45), EIAQDFK2ohibuTDLR (SEQ ID NO:
- 46), VTIMPK2ohibuDIQLAR (SEQ ID NO:
- 47), DNIQGITK2ohibuPAIR (SEQ ID NO:
- 48), K2ohibu TVTAMDVVYALK (SEQ ID NO:
- 49), TVTAMDVVYALK2ohibuR (SEQ ID NO:
- 50), KpropK2ohibuAAKpropKpropPAGVR (SEQ ID NO:
- 51), KpropKpropAAK2ohibuKpropPAGVR (SEQ ID NO:
- 51), KpropAAKpropK2ohibuPAGVR (SEQ ID NO:
- 52), K2ohibuASGPPVSELITKpropAVAASK2ohibu (SEQ ID NO:
- 53), KpropASGPPVSELITKpropAVAASK2ohibuER (SEQ ID NO:
- 54), ALAAAGYDVEK2ohibuNNSR (SEQ ID NO:
- 55), IK2ohibuLGLK (SEQ ID NO:
- 56), propGILVQTK2ohibuGTGASGSFKprop (SEQ ID NO:
- 57), propK2ohibuAASGEAKpropPQAKprop (SEQ ID NO:
- 58), propAASGEAK2ohibuPQAKprop(SEQ ID NO:
- 59), propAASGEAKpropPQAK2ohibuKprop (SEQ ID NO:
- 60), propAK2ohibuKpropPAGAAKprop (SEQ ID NO:
- 61), propAKpropKpropPAGAAK2ohibuKpropPKprop (SEQ ID NO:
- 62), propKpropATGAATPK2ohibuKprop (SEQ ID NO:
- 63), propKpropAK2ohibuKpropPAAAAVTKprop (SEQ ID NO:
- 64), propKpropPAAAAVTK2ohibuKprop (SEQ ID NO:
- 65), KpropVAAKpropKpropK2ohibu (SEQ ID NO:
- 66), GK2ohibuQGGK2ohibuAR (SEQ ID NO:
- 67), K2ohibuGNYSER (SEQ ID NO:
- 68), DNK2ohibuK2ohibuTR (SEQ ID NO:
- 69), NDEELNK2ohibuLLGR (SEQ ID NO:
- 70), VTIAQGGVLPNIQAVLLPK2ohibuK (SEQ ID NO:
- 71), propPEPAK2ohibuSAPAPKprop (SEQ ID NO:
- 72), PEPAK2ohibuSAPAPK (SEQ ID NO:
- 72), propPEPAKpropSAPAPKpropK2ohibuGSK (SEQ ID NO:
- 73), KpropAISK2ohibuAQKprop (SEQ ID NO:
- 74), AVTKpropAQK2ohibuKpropDGKpropKpropR (SEQ ID NO:
- 75), AVTKpropAQKpropK2ohibuDGKpropKpropR (SEQ ID NO:
- 75), K2ohibuESYSVYVYK (SEQ ID NO:
- 76), KESYSVYVYK2ohibuVLK (SEQ ID NO:
- 77), QVHPDTGISSK2ohibuAMGIMNSFVNDIFER (SEQ ID NO:
- 78), TK2ohibuQTAR (SEQ ID NO:
- 79), K2ohibuSTGGKacAPR (SEQ ID NO:
- 80), KpropSTGGK2ohibuAPR (SEQ ID NO:
- 80), K2ohibuQLATKacAAR (SEQ ID NO:
- 81), KQLATK2ohibuAAR (SEQ ID NO:
- 81), KacQLATK2ohibuAAR (SEQ ID NO:
- 81), K2ohibuSAPATGGVKpropKpropPHR (SEQ ID NO:
- 82), KpropSAPATGGVK2ohibuKpropPHR (SEQ ID NO:
- 82), K2ohibu LPFQR (SEQ ID NO:
- 83), K2ohibuGGKacGLGKacGGAKacR (SEQ ID NO:
- 84), GKacGGK2ohibuGLGKacGGAKacR (SEQ ID NO:
- 85), GLGK2ohibuGGAKacR (SEQ ID NO:
- 86), GGKpropGLGKpropGGAK2ohibuR (SEQ ID NO:
- 87), GGK2ohibuGLGKacGGAKacR (SEQ ID NO:
- 87), GGKacGLGK2ohibuGGAKacR (SEQ ID NO:
- 87), RGGVK2ohibuR (SEQ ID NO:
- 88), GVLK2ohibuVFLENVIR (SEQ ID NO:
- 89), KTVTAMDVVYALK2ohibuR (SEQ ID NO:
- 90), AGSAAK2ohibuASQSR (SEQ ID NO:
- 91), K2ohibuETYSSYIYK (SEQ ID NO:
- 92), ETYSSYIYK2ohibuVLK (SEQ ID NO:
- 93), IATEASK2ohibuLAAYNIK (SEQ ID NO:
- 94), LILPGELAK2ohibuHAVSEGTR (SEQ ID NO:
- 95), FQK2ohibuSTELLIR (SEQ ID NO:
- 96), DSVTYTEHAK2ohibuR (SEQ ID NO:
- 97), K2ohibuTVTSLDVVYALK (SEQ ID NO:
- 98), TVTSLDVVYALK2ohibuR (SEQ ID NO:
- 99), GMGK2ohibuVGAK (SEQ ID NO:
- 100), K2ohibuSAPATGGIK (SEQ ID NO:
- 101), YQK2ohibuSTDLLIR (SEQ ID NO:
- 102), VTIMTK2ohibuDMQLAR (SEQ ID NO:
- 103), K2ohibuAPAAAAEK (SEQ ID NO:
- 104), KacAPAAAAEK2ohibuK (SEQ ID NO:
- 105), VLK2ohibuQVHPDVGISK (SEQ ID NO:
- 106), IALESSK2ohibuLVR (SEQ ID NO:
- 107), HAISEGTK2ohibuAVTK (SEQ ID NO:
- 108), AVTK2ohibuFSSSTN (SEQ ID NO:
- 109), TASSK2ohibuQVSR (SEQ ID NO:
- 110), AGGKacAGK2ohibuDSGK (SEQ ID NO:
- 111), AGGK2ohibuAGKacDSGK (SEQ ID NO:
- 111) and SAPAPK2ohibuKacGSK (SEQ ID NO:
- 112), wherein K2ohibu is a 2-hydroxyisobutyrylated lysine, Kprop is a propanoylated lysine, and Kac is an acetylated lysine, wherein the isolated affinity reagent is an anti-histone H4 lysine 8 2-hydroxyisobutyrylation (H4K82ohibu) antibody, whereby the isolated affinity reagent and the histone protein or a fragment thereof form a binding complex;
- and
- (b) detecting the binding complex.
95 more elements
Dependencies are inferred from the claim text.