Protein farnesyltransferase and protein prenylation in Plasmodium falciparum.
Comparison of the malaria parasite and mammalian protein prenyltransferases and their cellular substrates is important for establishing this enzyme as a target for developing antimalarial agents. Nineteen heptapeptides differing only in their carboxyl-terminal amino acid were tested as alternative substrates of partially purified Plasmodium falciparum protein farnesyltransferase. Only NRSCAIM and NRSCAIQ serve as substrates, with NRSCAIM being the best. Peptidomimetics, FTI-276 and GGTI-287, inhibit the transferase with IC(50) values of 1 and 32 nm, respectively. Incubation of P. falciparum-infected erythrocytes with [(3)H]farnesol labels 50- and 22-28-kDa proteins, whereas [(3)H]geranylgeraniol labels only 22-28-kDa proteins. The 50-kDa protein is shown to be farnesylated, whereas the 22-28-kDa proteins are geranylgeranylated, irrespective of the labeling prenol. Protein labeling is inhibited more than 50% by either 5 microm FTI-277 or GGTI-298. The same concentration of inhibitors also inhibits parasite growth from the ring stage by 50%, decreases expression of prenylated proteins as measured with prenyl-specific antibody, and inhibits parasite differentiation beyond the trophozoite stage. Furthermore, differentiation specific prenylation of P. falciparum proteins is demonstrated. Protein labeling is detected predominantly during the trophozoite to schizont and schizont to ring transitions. These results demonstrate unique properties of protein prenylation in P. falciparum: a limited specificity of the farnesyltransferase for peptide substrates compared with mammalian enzymes, the ability to use farnesol to label both farnesyl and geranylgeranyl moieties on proteins, differentiation specific protein prenylation, and the ability of peptidomimetic prenyltransferase inhibitors to block parasite differentiation.
Department of Molecular Biology and Microbiology, University of Central Florida, Orlando 32816, USA.
Debopam
D
Chakrabarti
Thiago
T
Da Silva
Jennifer
J
Barger
Steve
S
Paquette
Hetal
H
Patel
Shelley
S
Patterson
Charles M
CM
Allen
D019883:Alkyl and Aryl Transferases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D000962:Antimalarials / Q000494:pharmacology; D004224:Diterpenes / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005204:Farnesol / Q000378:metabolism; D051231:Farnesyltranstransferase; D010963:Plasmodium falciparum / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism*; D017368:Protein Prenylation; D015800:Protozoan Proteins / Q000378:metabolism*; D013379:Substrate Specificity
D000962:Antimalarials; D004224:Diterpenes; D004791:Enzyme Inhibitors; D015800:Protozoan Proteins; D005204:Farnesol; C017338:geranylgeraniol; D019883:Alkyl and Aryl Transferases; D051231:Farnesyltranstransferase