HOW POTASSIUM AFFECTS THE ACTIVITY OF THE MOLECULAR CHAPERONE HSC70 .2. POTASSIUM BINDS SPECIFICALLY IN THE ATPASE ACTIVE-SITE

Crystallographic anomalous scattering from potassium at 1.7 Angstrom r esolution reveals two monovalent ions that interact with MgADP and P-i in the nucleotide binding cleft of wild-type recombinant bovine Hsc70 ATPase fragment. K+ at site 1 interacts with oxygens of the beta-phos phate of ADP, whereas K+ at site 2 interacts with an oxygen of P-i. Bo th K+ ions also interact with specific H2O molecules in the first hydr ation shell of the octahedrally coordinated Mg2+ ion and with specific protein ligands. In crystals that have Na+ present, K+ is replaced by a Na+ ion at site 1 and by a Na(+)similar to H2O pair at site 2. The K+ ions are positioned where they could stabilize binding of a beta,ga mma-bidentate MgATP complex with Hsc70, as well as a transition state during ATP hydrolysis, suggesting that monovalent ions act as specific metal cofactors in the ATPase reaction of Hsc70.