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Kinetoplastid membrane protein 11 induces pores in anionic phospholipid membranes: Effect of cholesterol.

Kinetoplastid membrane protein-11 (KMP-11), expressed in all stages of leishmanial life circle, is considered a potential candidate for leishmaniasis vaccine. KMP-11 is found on the membrane surface of the parasite. Although the biological function of KMP-11 is unknown, we hypothesize from its sequence analysis that it may interact with the macrophage membrane, and may influence the entry process of the parasite into the host cell. To validate this hypothesis, we have investigated the interaction of KMP-11 with the unilamellar anionic phospholipid vesicles and explored its pore forming activity. The decrease in negative zeta potential of the vesicles and reduction in fluorescence intensity of membrane bound dye DiI C-18 suggest a strong association of KMP-11 with membrane. The fluorescence leakage experiment as well as phase contrast microscopy show direct evidence of KMP-11induced pore formation in anionic membrane. Incorporation of cholesterol into membrane has been found to inhibit pore formation induced by KMP-11, suggesting an important role of cholesterol in leishmaniasis. Interestingly, vesicles containing only neutral phospholipid do not exhibit any tendency towards pore formation.