Bisphosphonates are potent inhibitors of Trypanosoma cruzi farnesyl pyrophosphate synthase.
We report the cloning and sequencing of a gene encoding the farnesyl pyrophosphate synthase of Trypanosoma cruzi. The protein (T. cruzi farnesyl pyrophosphate synthase, TcFPPS) is an attractive target for drug development, since the growth of T. cruzi is inhibited by carbocation transition state/reactive intermediate analogs of its substrates, the nitrogen-containing bisphosphonates currently in use in bone resorption therapy. The protein predicted from the nucleotide sequence of the gene has 362 amino acids and a molecular mass of 41.2 kDa. Several sequence motifs found in other FPPSs are present in TcFPPS. Heterologous expression of TcFPPS in Escherichia coli produced a functional enzyme that was inhibited by the nitrogen-containing bisphosphonates alendronate, pamidronate, homorisedronate, and risedronate but was less sensitive to the non-nitrogen-containing bisphosphonate etidronate, which, unlike the nitrogen-containing bisphosphonates, does not affect parasite growth. The protein contains a unique 11-mer insertion located near the active site, together with other sequence differences that may facilitate the development of novel anti-Chagasic agents.
Montalvetti
A
A
Laboratory of Molecular Parasitology, Department of Pathobiology, University of Illinois at Urbana-Champaign, Urbana, Illinois 61802, USA.
Bailey
B N
BN
Martin
M B
MB
Severin
G W
GW
Oldfield
E
E
Docampo
R
R
D019883:Alkyl and Aryl Transferases / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000235:genetics; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D001665:Binding Sites; D001717:Birds; D015152:Blotting, Northern; D015139:Blotting, Southern; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002412:Cations; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D004164:Diphosphonates / Q000737:chemistry*; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000378:metabolism; D012968:Etidronic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D051228:Geranyltranstransferase; D006863:Hydrogen-Ion Concentration; D008956:Models, Chemical; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011106:Polyisoprenyl Phosphates / Q000737:chemistry; D011485:Protein Binding; D011994:Recombinant Proteins / Q000378:metabolism; D000068296:Risedronic Acid; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012717:Sesquiterpenes; D014349:Trypanosoma cruzi / Q000201:enzymology*
D000596:Amino Acids; D002121:Calcium Channel Blockers; D002412:Cations; D004164:Diphosphonates; D011106:Polyisoprenyl Phosphates; D011994:Recombinant Proteins; D012717:Sesquiterpenes; C015234:geranyl pyrophosphate; C004808:farnesyl pyrophosphate; D019883:Alkyl and Aryl Transferases; D051228:Geranyltranstransferase; D000068296:Risedronic Acid; D012968:Etidronic Acid