CPYA 4.2769 853# ' X,eXW@ P??flowdialysis_AP h㈵?9̗?-C6 ?K8 ?jUMu?D8 ?מY?b2U0*?h㈵?q@H?9̗?-C6?L8?iUMu?D8 !?מY"?b2U0*#?h㈵$?q@H&?9̗'? W(?-C6*?ZӼ? `??flowdialysis_BP 333333I@33333Z@y[@ \@fffff\@ ^@`_@@`@\a@a@fffffc@fffffc@f@fffffj@fffffn@r@̌u@y@33333O@@H@Ĕ@@j@ flowdialysis ^DLPOrigin,FdXXQ.DBAQ.DBAXX - ^@?@@@?9@l "$$] (? ף= ף= Pd, o #c5fffff?Կ(I>>__WIOTN (  Ifffff?Կ(I I o92 o #5Ifffff??(>>__LayerInfoStorage7 (  Ifff??(  AP$$  total [Ca2+] (M)  B`2$  cpm in eluate    P  @ P   "&@%3% P  @$2$ P    P   P P    P  @ P   "&@%%V P  @$$U P    P   P P   P   P   P   P   P   P @ VTT t.DBA BA  Notes  The data in the "flow_dialysis" spreadsheet describe the binding of Ca2+ to a mutant of rat b-parvalbumin. The protein concentration was 104 M. a. Obtain estimates for the site-specific binding constants. b. Obtain estimates for the stepwise macroscopic constants. c. Are the values of the macroscopic and microscopic constants in agreement with prediction? d. Prepare a logarithmic Hill plot for the system. Comment on its appearance. e. Generate species distribution curves for the apo-, singly-, and doubly-bound species. f. Calculate the median ligand concentration. What is the fraction of singly-bound protein at that concentration? g. Calculate the overall free energy change for the reaction. @ ResultsLog    6 )gP.DBA}'BA  flow-dialysis_2008            %  (