Abstract
The membrane-bound protein family, P-type ATPases, couples ATP hydrolysis with substrate transport across the membrane and forms an obligatory auto-phosphorylated intermediate in the transport cycle. The metal fluoride compounds, BeF x , AlF x , and MgF x , as phosphate analogs stabilize different enzyme structural states in the phosphoryl transfer/hydrolysis reactions, thereby fixing otherwise short-lived intermediate and transient structural states and enabling their biochemical and atomic-level crystallographic studies. The compounds thus make an essential contribution for understanding of the ATP-driven transport mechanism. Here, with a representative member of P-type ATPase, sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA), we describe the method for their binding and for structural and functional characterization of the bound states, and their assignments to states occurring in the transport cycle.
Published Version
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