Journal: Science Advances
Article Title: Molecular basis of hUHRF1 allosteric activation for synergistic histone modification binding by PI5P
doi: 10.1126/sciadv.abl9461
Figure Lengend Snippet: ( A to F ) Titration series of H3K9me3 peptide (in the absence and presence of di-C 16:0 PI5P), PI5P, or PI3P with fluorescently labeled, mutant, and wild-type, full-length hUHRF1 were analyzed by MST. PBR*: K644A, K646A, K648A, R649A, K650A, S651A; R296*: R296A; R649*: R649A; D142*: D142A; R296R649*: R296A, R649A; and D142R649*: D142A, R649A. n = 3; error bars: SD; K d values are listed in tables S1 and S2.
Article Snippet: Phosphatidylinositol 5-phosphate diC16 (di-C 16:0 PI5P; Echelon, #P5016), phosphatidylinositol 4-phosphate diC16 (di-C 16:0 PI4P; Echelon, #P4016), phosphatidylinositol 3-phosphate diC16 (di-C 16:0 PI3P; Echelon, #P3016), phosphatidylinositol 4,5-bisphosphate diC16 [di-C 16:0 PI(4,5)P2; Echelon, #P4516], phosphatidylinositol 3,5-bisphosphate diC16 [di-C 16:0 PI(3,5)P2; Echelon, #P3516], phosphatidylinositol 5-phosphate diC8 (di-C 8:0 PI5P; Echelon, #P-5008), inositol 1,5-bisphosphate [Ins(1,5)P2; Echelon, #Q-0015], 1,2-dioleoyl- sn -glycero-3-phospho-(1′-myo-inositol-5′-phosphate) (di-C 18:1 PI5P; Avanti Polar Lipids, #850152P), 1-heptadecanoyl-2-(5Z,8Z,11Z,14Z-eicosatetraenoyl)- sn -glycero-3-phospho-(1′-myo-inositol-5′-phosphate) (17:0,20:4 PI5P; Avanti Polar Lipids, #LM1902), 1,2-dipalmitoyl- sn -glycero-3-phosphoethanolamine- N -(biotinyl) (sodium salt) (di-C 16:0 biotinyl PE; Avanti Polar Lipids, #870285), 1,2-dimyristoyl- sn -glycero-3-phosphoethanolamine (di-C 14:0 PE; Avanti Polar Lipids, #850745), 1,2-dioleoyl- sn -glycero-3-phosphoethanolamine- N -(biotinyl) (sodium salt) (di-C 18:1 biotinyl PE; Avanti Polar Lipids, #870282), 1,2-distearoyl- sn -glycero-3-phosphoethanolamine- N -[methoxy(polyethylene glycol)-550] (ammonium salt) (di-C 18:0 PEG550PE; Avanti Polar Lipids, #880520), cholesterol-(polyethylene glycol-600) (Avanti Polar Lipids, #880001), 1,2-dipalmitoyl- sn -glycero-3-phosphate (di-C 16:0 PA; Avanti Polar Lipids, #830855), 1,2-dimyristoyl- sn -glycero-3-phosphocholine (DMPC) (Anatrace, #D514), and 1,2-diheptanoyl- sn -glycero-3-phosphocholine (DHPC) (Anatrace, #D607).
Techniques: Titration, Labeling, Mutagenesis