Background/Aims: In a previous study, me have demonstrated that primary human hepatocytes in culture are susceptible for Plasmodium falciparum sporozoite invasion and for development of parasites into exoerythrocytic forms. In a separate study we demonstrated the involvement of two human liver plasma membrane proteins (55 kD and 20 kD) in the invasion of P. falciparum sporozoites in vitro. In this study, we have unravelled the nature of the 55 kD protein.
Methods: For the identification of this protein, a 53-58 kD membrane protein fraction from human liver was isolated, radioactively labelled, incubated with sporozoites and cross-linked, After reduction of the cross-linker, the released proteins were mixed with unlabelled 53-58 kD protein fraction and separated on two-dimensional SDS-PAGE, Autoradiography showed a single spot corresponding to a protein of 55 kD and pI of 5.7-5.8.
Results: Amino acid sequencing revealed the 55 kD protein as carboxylesterase, The biological activity of purified human liver carboxylesterase and of an anti-serum against carboxylesterase on sporozoite invasion in vitro was evaluated, Human carboxylesterase as well as a rabbit antiserum against carboxylesterase inhibited the invasion of P. falciparum sporozoites into primary human hepatocytes in culture, A number of carboxylesterase cDNA clones were isolated from a human liver cDNA library, Sequence analysis revealed two different iso-types, Immunoaffinity purified recombinant human carboxylesterase was shown also to inhibit the invasion of sporozoites into primary human hepatocytes, Immunocytochemical analysis of the localisation of carboxylesterase in primary cultures of human hepatocytes using specific antibodies, showed its presence inside the hepatocytes and on the membrane.
Conclusions: Carboxylesterase plays a role in the invasion process of P. falciparum sporozoites into human hepatocytes in vitro. The implications of these findings are further discussed.
|Number of pages||11|
|Journal||Journal of Hepatology|
|Publication status||Published - Oct-1997|
- amino acid sequence
- liver stage parasites
- parasitic disease
- CIRCUMSPOROZOITE PROTEIN
- MEMBRANE PROTEINS