Figure 4
(A) Alignments of the aa sequences of the PdVP1 and PdVP2 isoenzymes of H+-PPases of P. dicentrarchi using the Blastp program (https://blast.ncbi.nlm.nih.gov/Blast.cgi). The blue shaded sequence corresponds to the fragment of the aa sequence of the PdVP1 isoenzyme expressed as a recombinant protein in the yeast Klyuveromyces lactis (rPdVP1), and the green shaded sequence corresponds to the HKAAVIGDTIGDPLK (HK) peptide of the isoenzyme PdVP1. The alignments of the PABHK motif in the V-H+-PPases AVP1 and AVP2 of Arabidopsis thaliana and the V-H+-PPases PdVP1 and PdVP2 of P. dicentrarchi, shown in pink shading, represent the common regions. (B) Western blot analysis of components of a P. dicentrarchi ciliate lysate (CL) separated by SDS-PAGE under non-reducing (-DTT; lanes 1 and 3) and reducing (+ DTT; lanes 2 and 4) conditions in the presence of polyclonal mouse antibodies α-rPdVP1 (lanes 1 and 2) and α-KLH-HK (lanes 3 and 4). Mw: molecular weight markers in kDa. (C,D) Immunolocalization of H+-PPases of P. dicentrarchi by immunofluorescence by use of polyclonal antibodies α-rPdVP1(C) and α-KLH-HK (D) with arrows indicating labelling sites.
