The HslV Protease from Leishmania major and Its Activation by C-terminal HslU Peptides
AbstractHslVU is an ATP-dependent proteolytic complex present in certain bacteria and in the mitochondrion of some primordial eukaryotes, including deadly parasites such as Leishmania. It is formed by the dodecameric protease HslV and the hexameric ATPase HslU, which binds via the C-terminal end of its subunits to HslV and activates it by a yet unclear allosteric mechanism. We undertook the characterization of HslV from Leishmania major (LmHslV), a trypanosomatid that expresses two isoforms for HslU, LmHslU1 and LmHslU2. Using a novel and sensitive peptide substrate, we found that LmHslV can be activated by peptides derived from the C-termini of both LmHslU1 and LmHslU2. Truncations, Ala- and D-scans of the C-terminal dodecapeptide of LmHslU2 (LmC12-U2) showed that five out of the six C-terminal residues of LmHslU2 are essential for binding to and activating HslV. Peptide cyclisation with a lactam bridge allowed shortening of the peptide without loss of potency. Finally, we found that dodecapeptides derived from HslU of other parasites and bacteria are able to activate LmHslV with similar or even higher efficiency. Importantly, using electron microscopy approaches, we observed that the activation of LmHslV was accompanied by a large conformational remodeling, which represents a yet unidentified layer of control of HslV activation. View Full-Text
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Kebe, N.M.; Samanta, K.; Singh, P.; Lai-Kee-Him, J.; Apicella, V.; Payrot, N.; Lauraire, N.; Legrand, B.; Lisowski, V.; Mbang-Benet, D.-E.; Pages, M.; Bastien, P.; Kajava, A.V.; Bron, P.; Hernandez, J.-F.; Coux, O. The HslV Protease from Leishmania major and Its Activation by C-terminal HslU Peptides. Int. J. Mol. Sci. 2019, 20, 1021.
Kebe NM, Samanta K, Singh P, Lai-Kee-Him J, Apicella V, Payrot N, Lauraire N, Legrand B, Lisowski V, Mbang-Benet D-E, Pages M, Bastien P, Kajava AV, Bron P, Hernandez J-F, Coux O. The HslV Protease from Leishmania major and Its Activation by C-terminal HslU Peptides. International Journal of Molecular Sciences. 2019; 20(5):1021.Chicago/Turabian Style
Kebe, Ndeye M.; Samanta, Krishnananda; Singh, Priyanka; Lai-Kee-Him, Joséphine; Apicella, Viviana; Payrot, Nadine; Lauraire, Noémie; Legrand, Baptiste; Lisowski, Vincent; Mbang-Benet, Diane-Ethna; Pages, Michel; Bastien, Patrick; Kajava, Andrey V.; Bron, Patrick; Hernandez, Jean-François; Coux, Olivier. 2019. "The HslV Protease from Leishmania major and Its Activation by C-terminal HslU Peptides." Int. J. Mol. Sci. 20, no. 5: 1021.
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