Skip to main content
Figure 4 | BMC Neuroscience

Figure 4

From: Targeted inhibition of the Shroom3–Rho kinase protein–protein interaction circumvents Nogo66 to promote axon outgrowth

Figure 4

A single cysteine residue in the Shroom3 SD2 domain is required for CCG-17444 activity. a CCG-17444 activity is blocked by DTT. To investigate a thiol sensitive mechanism, DTT was added after plating SD2 and was present at all subsequent steps in the ELISA. (n = 3, p < 0.001, Student’s t test). b CCG-17444 is an irreversible inhibitor. After plating, SD2 was treated with CCG-17444 then wells were washed extensively to remove unbound compound prior to addition of R2-C1 (n = 3). c Cys254 of SD2 is the site of action of CCG-17444. CCG-17444 inhibits the interaction of WT SD2 with the ROCK R2-C1 domain but does not inhibit the interaction of SD2 Cys254Ala with ROCK (upper left panel). WT and Cys254Ala SD2 interact to similar extent with ROCK (upper right panel). CCG-17444 inhibits the interaction of SD2 Cys379Ala with R2-C1, suggesting that Cys379A is not a site of drug action (lower left panel). The double Cys to Ala mutant is indistinguishable from the single Cys254Ala mutant (lower right panel), indicating a single site of action of CCG-17444 (n = 3).

Back to article page