Characterization of the lipid-binding domain of the Plasmodium falciparum CTP:phosphocholine cytidylyltransferase through synthetic-peptide studies
| Autor: | Claude Roustan, Henri Vial, Rachel Cerdan, Patrick Seta, Catherine Gumila, Luc Maurin, Marie-Pierre Larvor |
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| Přispěvatelé: | Dynamique des interactions membranaires normales et pathologiques (DIMNP), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université Montpellier 1 (UM1), Institut Européen des membranes (IEM), Centre National de la Recherche Scientifique (CNRS)-Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Institut de Chimie du CNRS (INC)-Université de Montpellier (UM), Dynamique moléculaire des interactions membranaires (DMIM), Centre National de la Recherche Scientifique (CNRS)-Université Montpellier 2 - Sciences et Techniques (UM2) |
| Jazyk: | angličtina |
| Rok vydání: | 2003 |
| Předmět: |
Protein Conformation
Surface Properties Plasmodium falciparum Cytidylyltransferase Peptide Biology Biochemistry Choline-phosphate cytidylyltransferase 03 medical and health sciences chemistry.chemical_compound Protein structure Animals [CHIM]Chemical Sciences Choline-Phosphate Cytidylyltransferase Lipid bilayer Molecular Biology ComputingMilieux_MISCELLANEOUS 030304 developmental biology Phosphocholine chemistry.chemical_classification 0303 health sciences Liposome Binding Sites Circular Dichroism 030302 biochemistry & molecular biology Water Cell Biology Phosphatidylserine Lipid Metabolism Kinetics Spectrometry Fluorescence chemistry Liposomes Adsorption Oligopeptides Protein Binding Research Article |
| Zdroj: | Biochemical Journal Biochemical Journal, Portland Press, 2003, 375 (3), pp.653-661. ⟨10.1042/BJ20031011⟩ |
| ISSN: | 0264-6021 1470-8728 |
| DOI: | 10.1042/BJ20031011⟩ |
| Popis: | Phospholipid biosynthesis plays a key role in malarial infection and is regulated by CCT (CTP:phosphocholine cytidylyltransferase). This enzyme belongs to the group of amphitropic proteins which are regulated by reversible membrane interaction. To assess the role of the putative membrane-binding domain of Plasmodium falciparum CCT (PfCCT), we synthesized three peptides, K21, V20 and K54 corresponding to residues 274–294, 308–327 and 274–327 of PfCCT respectively. Conformational behaviour of the peptides, their ability to bind to liposomes and to destabilize lipid bilayers, and their insertion properties were investigated by different biophysical techniques. The intercalation mechanisms of the peptides were refined further by using surface-pressure measurements on various monolayers at the air/water interface. In the present study, we show that the three studied peptides are able to bind to anionic and neutral phospholipids, and that they present an α-helical conformation upon lipid binding. Peptides V20 and the full-length K54 intercalate their hydrophobic parts into an anionic bilayer and, to a lesser extent, a neutral one for V20. Peptide K21 interacts only superficially with both types of phospholipid vesicles. Adsorption experiments performed at the air/water interface revealed that peptide K54 is strongly surface-active in the absence of lipid. Peptide V20 presents an atypical behaviour in the presence of phosphatidylserine. Whatever the initial surface pressure of a phosphatidylserine film, peptide V20 and phosphatidylserine entities seem linked together in a special organization involving electrostatic and hydrophobic interactions. We showed that PfCCT presents different lipid-dependence properties from other studied CCTs. Although the lipid-binding domain seems to be located in the C-terminal region of the enzyme, as with the mammalian counterpart, the membrane anchorage, which plays a key role in the enzyme regulation, is driven by two α-helices, which behave differently from one another. |
| Databáze: | OpenAIRE |
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