Characterization and immunolocalization of a main proteinaceous component of the cell wall of the protozoan parasite Perkinsus atlanticus

Abstract

Described in the present study is a major component of the cell wall of two of the most pathogenic parasites of molluscs, Perkinsus atlanticus and P. marinus. The component is a high-molecular-weight protein (233-kDa), which we have named PWP-1 (for Perkinsus wall protein-1). Western blots, using a polyclonal serum generated against purified PWP-1 from P. atlanticus, revealed that this protein is expressed by all walled developmental stages of this protozoon. By means of immunogold electron microscopy, labelling for PWP-1 was strong and specifically associated with the cell wall. The label density and distribution pattern was quite different between trophozoites and prezoosporangia. With regard to the structural organization of this protein, PWP-1 is disulphide-linked to other cell wall components and released from the cell wall only following treatment with a sulfhydryl agent. We also report that PWP-1 is a trypsin-resistant protein, both in its native and heat-denatured conformation. In addition, results from the N-terminal microsequence of this protein allow us to define PWP-1 as a novel cell wall protein. Overall, our findings strongly suggest that PWP-1 plays a key role in the organization of the cell wall of these protozoa promoting their survival.