Fig 5. The gCLCA1EQ mutant eliminated autocatalytic cleavage similar to CLCA proteins of mammalian cluster 1.

Immunoblotting of HEK293 cell lysates. The left panel illustrates immunoblots from murinized galline CLCA1, the central panel from murine CLCA1 and the right panel from murine CLCA4a constructs. The proteins of the three homologues differ in their molecular weights. The mutation of the HExxH motif of gCLCA1 eliminated the cleavage of the ~154 kDa precursor protein (*) similarly to the murine CLCA1 protein. In contrast, the cleavage of the murine CLCA4aEQ mutant was only impaired, but not totally absent. Asterisks (*) indicate the uncleaved protein of the respective CLCA homolog. To control for equal total protein loading, the samples were identically immunoblotted with primary anti-beta-actin antibodies. WT = HExxH wild type motif, EQ = EQ mutation in the HExxH motif, C = mock-transfected control. Representative images of three independent experiments.