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Crystal structure of soluble domain of malaria sporozoite protein UIS3 in complex with lipid.

Sharma A, Yogavel M, Akhouri RR, Gill J, Sharma A - J. Biol. Chem. (2008)

Bottom Line: We additionally provide new structural and biochemical evidence of PfUIS3(130-229) interactions with lipids (phosphatidylethanolamine), with phospholipid liposomes, and with the human liver fatty acid-binding protein.The direct interaction of PfUIS3(130-229) with liver fatty acid-binding protein most likely provides the parasite with a conduit for importing essential fatty acids/lipids.Therefore, our analyses have implications for lipid transport into the parasite during the rapid growth phases of sporozoites.

View Article: PubMed Central - PubMed

Affiliation: Structural and Computational Biology Group, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Road, New Delhi, India.

ABSTRACT
Malaria parasite UIS3 (up-regulated in infective sporozoites gene 3) is essential for sporozoite development in infected hepatocytes. UIS3 encodes for a membrane protein that is localized to the parasite parasitophorous vacuolar membrane in infected hepatocytes. We describe here 2.5-A resolution crystal structure of Plasmodium falciparum UIS3 soluble domain (PfUIS3(130-229)) in complex with the lipid phosphatidylethanolamine (PE). PfUIS3(130-229) is a novel, compact, and all alpha-helical structure bound to one molecule of PE. The PfUIS3(130-229)-PE complex structure reveals a novel binding site with specific interactions between PfUIS3(130-229) and the PE head group. One acyl chain of PE wraps around part of PfUIS3(130-229) and docks onto a hydrophobic channel. We additionally provide new structural and biochemical evidence of PfUIS3(130-229) interactions with lipids (phosphatidylethanolamine), with phospholipid liposomes, and with the human liver fatty acid-binding protein. The direct interaction of PfUIS3(130-229) with liver fatty acid-binding protein most likely provides the parasite with a conduit for importing essential fatty acids/lipids. Therefore, our analyses have implications for lipid transport into the parasite during the rapid growth phases of sporozoites. Given that PfUIS3 is essential for establishment of liver stage infection by P. falciparum, our data provide a new target for abrogating parasite development within liver cells before typical symptoms of malaria can manifest.

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Related in: MedlinePlus

Binding of PfUIS3130-229 to human LFABP. Protein-protein based ELISAs were performed as described under “Experimental Procedures.” Each experiment was performed in quadruplicate, and an average of four experiments was plotted after deducting the background signal from the negative control (Gelatin). The standard deviations were less than 10% in all cases. SDS-PAGE of purified PfUIS3 and LFABP used in these ELISAs are shown as insets in the figure.
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fig10: Binding of PfUIS3130-229 to human LFABP. Protein-protein based ELISAs were performed as described under “Experimental Procedures.” Each experiment was performed in quadruplicate, and an average of four experiments was plotted after deducting the background signal from the negative control (Gelatin). The standard deviations were less than 10% in all cases. SDS-PAGE of purified PfUIS3 and LFABP used in these ELISAs are shown as insets in the figure.

Mentions: PfUIS3130-229 Binds Human Liver Fatty Acid-binding Protein—The rodent malaria parasite homolog of PfUIS3, PyUIS3, is known to interact with mouse liver fatty acid-binding protein as shown previously by yeast two-hybrid screens and co-immunoprecipitation experiments (4). Not surprisingly, we found that the homolog of PyUIS3 in P. falciparum interacts with the human counterpart of rodent LFABP. Our evidence for PfUIS3130-229-human FABP interaction is based on purified components used in protein-protein based ELISAs (Fig. 10). The binding between PfUIS3130-229 and human LFABP was specific and dose-dependent. Further, this experiment clearly suggests that the recognition site for human LFABL on PfUIS3 resides within the monomeric 130-229 spanning domain.


Crystal structure of soluble domain of malaria sporozoite protein UIS3 in complex with lipid.

Sharma A, Yogavel M, Akhouri RR, Gill J, Sharma A - J. Biol. Chem. (2008)

Binding of PfUIS3130-229 to human LFABP. Protein-protein based ELISAs were performed as described under “Experimental Procedures.” Each experiment was performed in quadruplicate, and an average of four experiments was plotted after deducting the background signal from the negative control (Gelatin). The standard deviations were less than 10% in all cases. SDS-PAGE of purified PfUIS3 and LFABP used in these ELISAs are shown as insets in the figure.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2527117&req=5

fig10: Binding of PfUIS3130-229 to human LFABP. Protein-protein based ELISAs were performed as described under “Experimental Procedures.” Each experiment was performed in quadruplicate, and an average of four experiments was plotted after deducting the background signal from the negative control (Gelatin). The standard deviations were less than 10% in all cases. SDS-PAGE of purified PfUIS3 and LFABP used in these ELISAs are shown as insets in the figure.
Mentions: PfUIS3130-229 Binds Human Liver Fatty Acid-binding Protein—The rodent malaria parasite homolog of PfUIS3, PyUIS3, is known to interact with mouse liver fatty acid-binding protein as shown previously by yeast two-hybrid screens and co-immunoprecipitation experiments (4). Not surprisingly, we found that the homolog of PyUIS3 in P. falciparum interacts with the human counterpart of rodent LFABP. Our evidence for PfUIS3130-229-human FABP interaction is based on purified components used in protein-protein based ELISAs (Fig. 10). The binding between PfUIS3130-229 and human LFABP was specific and dose-dependent. Further, this experiment clearly suggests that the recognition site for human LFABL on PfUIS3 resides within the monomeric 130-229 spanning domain.

Bottom Line: We additionally provide new structural and biochemical evidence of PfUIS3(130-229) interactions with lipids (phosphatidylethanolamine), with phospholipid liposomes, and with the human liver fatty acid-binding protein.The direct interaction of PfUIS3(130-229) with liver fatty acid-binding protein most likely provides the parasite with a conduit for importing essential fatty acids/lipids.Therefore, our analyses have implications for lipid transport into the parasite during the rapid growth phases of sporozoites.

View Article: PubMed Central - PubMed

Affiliation: Structural and Computational Biology Group, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Road, New Delhi, India.

ABSTRACT
Malaria parasite UIS3 (up-regulated in infective sporozoites gene 3) is essential for sporozoite development in infected hepatocytes. UIS3 encodes for a membrane protein that is localized to the parasite parasitophorous vacuolar membrane in infected hepatocytes. We describe here 2.5-A resolution crystal structure of Plasmodium falciparum UIS3 soluble domain (PfUIS3(130-229)) in complex with the lipid phosphatidylethanolamine (PE). PfUIS3(130-229) is a novel, compact, and all alpha-helical structure bound to one molecule of PE. The PfUIS3(130-229)-PE complex structure reveals a novel binding site with specific interactions between PfUIS3(130-229) and the PE head group. One acyl chain of PE wraps around part of PfUIS3(130-229) and docks onto a hydrophobic channel. We additionally provide new structural and biochemical evidence of PfUIS3(130-229) interactions with lipids (phosphatidylethanolamine), with phospholipid liposomes, and with the human liver fatty acid-binding protein. The direct interaction of PfUIS3(130-229) with liver fatty acid-binding protein most likely provides the parasite with a conduit for importing essential fatty acids/lipids. Therefore, our analyses have implications for lipid transport into the parasite during the rapid growth phases of sporozoites. Given that PfUIS3 is essential for establishment of liver stage infection by P. falciparum, our data provide a new target for abrogating parasite development within liver cells before typical symptoms of malaria can manifest.

Show MeSH
Related in: MedlinePlus