Figure 6.

In vitro binding assay identifying the cain/cabin1-binding domain in the Cn A subunit. (A) Schematic diagram showing deletion mutants of Cn A (CnA): The CnAΔCaMAI mutant lacks the calmodulin (CaM)-binding and autoinhibitory (AI) domains and the CnAΔeD mutant lacks Cn B (CnB)-binding region and contains only the catalytic domain. (B) Purified GST and GST-cain/cabin1-C were subjected to in vitro binding assays by using [³⁵S]methionine-labeled full-length or deletions of Cn A. Interacting proteins bound to GST-cain/cabin1-C were pulled down with GST-Sepharose, separated by SDS/PAGE, and exposed to x-ray film for autoradiography.