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. Author manuscript; available in PMC: 2019 Feb 9.
Published in final edited form as: Biochemistry. 2016 May 4;55(19):2794–2805. doi: 10.1021/acs.biochem.6b00006

Figure 7.

Figure 7.

Effect of R/K point mutations on the interaction of Purβ with MSY1. (A) The binding of purified Purβ wild-type (WT) protein or the indicated single (R267A), double (R159A/R267A), or triple (K82A/R159A/R267A) point mutants to immobilized MSY1 (filled symbols, solid lines) or BSA (open symbols, dashed lines) was evaluated by ELISA. Purβ-MSY1 complexes were detected using rabbit anti-Purβ 210–229 as the primary antibody. Binding isotherms were generated by fitting data points obtained from several independent titration experiments (n = 3) to the equation for a rectangular hyperbola. (B) Binding curves generated after subtracting out the nonspecific absorbance measured in BSA only-coated wells from the absorbance obtained in MSY1-coated wells.