Detailed characterization of a cyclophilin from the human malaria parasite Plasmodium falciparum

Matthew Berriman, Alan H. Fairlamb (Lead / Corresponding author)

    Research output: Contribution to journalArticlepeer-review

    45 Citations (Scopus)

    Abstract

    Cyclosporin (Cs)A has pronounced antimalarial activity in vitro and in vivo. In other organisms, the drug is thought to exert its effects either by inhibiting the peptidylprolyl cis/trans isomerase activity of cyclophilin (CyP) or by forming a CyP-CsA complex that inhibits the phosphatase activity of calcineurin. We have cloned and overexpresscd in Escherichia coli a gene encoding a CyP from Plasmodium falciparum (PfCyP19) that is located on chromosome 3. The sequence of PfCyP19 shows remarkable sequence identity with human CyPA and, unlike the two previously identified CyPs from P. falciparum, PfCyP19 has no signal peptide or N-terminal sequence extension, suggesting a cytosolic localization. All the residues implicated in the recognition of the synthetic substrate N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide are conserved, resulting in characteristically high Michaelis-Menten and specificity constants (Km >> 120 μM, kcat/Km = 1.2 x 107 M-1̇s-1 respectively). As the first line in the functional characterization of this enzyme, we have assessed its binding affinity for CsA. In accordance with its tryptophan-containing CsA-binding domain, PfCyP19 binds CsA with high affinity (Kd = 13 nM, Ki = 6.9 nM). Twelve CsA analogues were also found to possess Ki values similar to CsA, with the notable exceptions of Val2-Cs (Ki = 218 nM) and Thr2-Cs (Ki = 690 nM). The immunosuppressants rapamycin and FK506 were inactive as inhibitors, consistent with other members of the CyP family of rotamases. The CsA analogues were also assessed as inhibitors of P. falciparum growth in vitro. Although their antimalarial activity did not correlate with inhibition of enzyme activity, residues 3 and 4 of CsA appeared to be important for inhibition of parasite growth and residues 1 and 2 for PfCyP19 inhibition. We propose that a malarial CyP-CsA complex presents residues 3 and 4 as part of an 'effector surface' for recognition by a downstream target, similar to the proposed mechanism for T-cell immunosuppression.

    Original languageEnglish
    Pages (from-to)437-445
    Number of pages9
    JournalBiochemical Journal
    Volume334
    Issue number2
    DOIs
    Publication statusPublished - 1 Sep 1998

    Fingerprint

    Dive into the research topics of 'Detailed characterization of a cyclophilin from the human malaria parasite Plasmodium falciparum'. Together they form a unique fingerprint.

    Cite this