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. 2020 Jan 20;295(9):2866–2884. doi: 10.1074/jbc.RA119.010077

Figure 1.

Figure 1.

A, bar charts showing the relative enrichment of peptides in each library pool and the 10 most frequently selected sequences under each set of conditions. Sequences taken forward for further evaluation are highlighted in red. B, ELISA screening of phage presenting the three most enriched sequences. The relative absorbances from ELISA for binding against a panel of small GTPase targets are as indicated, and a BSA control is plotted as a modified box plot. n = 3–4. C, displacement of [3H]GTP·Cdc42 from GST–WT ACK GBD by C1 binding to Cdc42. Increasing concentrations of C1 or C1C4S, C13S were titrated into a fixed concentrations of [3H]GTP·Cdc42 and GST–WT ACK GBD in the presence or absence of 50 mm DTT, in competition SPAs. The Kd value for Cdc42 binding to GST–WT ACK GBD was fixed to the value (30 nm) obtained for this Kd in direct SPAs. The data were fitted to an isotherm describing a pure competition model (58) to give an apparent Kd (Ki) value for C1. The data and curve fits are displayed as a percentage of the maximal SPA signal in each assay.