pH titration studies of an SH2 domain-phosphopeptide complex: unusual histidine and phosphate pKa values.
Electrostatic interactions in a complex of the phospholipase C-gamma 1 C-terminal SH2 domain with a high-affinity binding phosphopeptide representing the sequence around Tyr 1021 of the beta platelet-derived growth factor receptor were studied by pKa determination of various titratable groups over the pH range of 5 to 8. A histidine residue that is highly conserved among SH2 domains (His beta D4) and the phosphotyrosine (pTyr) phosphate group show pKa values significantly lower than average for these residue types in proteins. The reduced pKa of these two groups is due to the proximity of the highly positively charged pTyr binding pocket. The unusual pKa of His beta D4 is also due to burial from solvent in a hydrogen-bonding network that appears necessary for the positioning of arginine residues involved in pTyr binding. Mutation of the analogous histidine in other SH2 domains has been shown to abrogate pTyr binding. In addition to these large shifts in pKa values, smaller effects were observed for the titratable groups of a glutamic acid and histidine near the C-terminus of the the second helix due to its helical dipole. Finally, exchange behavior of arginine guanidinium protons with solvent as a function of pH in this SH2 domain-phosphopeptide complex confirms previous descriptions of the roles of different arginines in the structure and function of this protein.
Singer
A U
AU
Department of Molecular and Medical Genetics, University of Toronto, Canada.
Forman-Kay
J D
JD
D000577:Amides / Q000737:chemistry; D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D018698:Glutamic Acid / Q000737:chemistry; D006639:Histidine / Q000737:chemistry*; D006860:Hydrogen Bonding; D006863:Hydrogen-Ion Concentration; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D010446:Peptide Fragments / Q000737:chemistry*; D010710:Phosphates / Q000737:chemistry*; D010750:Phosphoproteins / Q000737:chemistry*; D019000:Phosphotyrosine / Q000737:chemistry; D017433:Protein Structure, Secondary; D020797:Receptor, Platelet-Derived Growth Factor beta; D017479:Receptors, Platelet-Derived Growth Factor / Q000737:chemistry*; D013438:Sulfhydryl Compounds / Q000737:chemistry; D010738:Type C Phospholipases / Q000737:chemistry*
D000577:Amides; D010446:Peptide Fragments; D010710:Phosphates; D010750:Phosphoproteins; D013438:Sulfhydryl Compounds; D019000:Phosphotyrosine; D018698:Glutamic Acid; D006639:Histidine; D020797:Receptor, Platelet-Derived Growth Factor beta; D017479:Receptors, Platelet-Derived Growth Factor; D010738:Type C Phospholipases