Identification and deletion of the PEST-like sequences in the CaVβ3 subunit. A, Schematic representation of the functional domains of CaVβ3 and the putative PEST sequences. Two potential PEST regions with scores of +9.47 (residues 24–37) and +11.52 (residues 397–411) were found in the amino acid sequence of CaVβ3 when analyzed with the PESTFind software. SH3 denotes a type 3 src-homology and GK-like indicate a guanylate kinase domain. B, Autoradiogram of in vitro translated [35S]-methionine-labeled wild-type (β3) and PEST deletion mutants of the CaVβ3 subunit (ΔP1, ΔP2, ΔP1–2) resolved by SDS-PAGE. 5 μl of each translation reaction were run per lane. C, Western blot analysis of membranes from untransfected HEK-293 cells (−) or cells expressing the wild-type (β3) and the CaVβ3 PEST deletion mutants (ΔP1, ΔP2, ΔP1–2).