Squalene synthase as a target for Chagas disease therapeutics.
Trypanosomatid parasites are the causative agents of many neglected tropical diseases and there is currently considerable interest in targeting endogenous sterol biosynthesis in these organisms as a route to the development of novel anti-infective drugs. Here, we report the first x-ray crystallographic structures of the enzyme squalene synthase (SQS) from a trypanosomatid parasite, Trypanosoma cruzi, the causative agent of Chagas disease. We obtained five structures of T. cruzi SQS and eight structures of human SQS with four classes of inhibitors: the substrate-analog S-thiolo-farnesyl diphosphate, the quinuclidines E5700 and ER119884, several lipophilic bisphosphonates, and the thiocyanate WC-9, with the structures of the two very potent quinuclidines suggesting strategies for selective inhibitor development. We also show that the lipophilic bisphosphonates have low nM activity against T. cruzi and inhibit endogenous sterol biosynthesis and that E5700 acts synergistically with the azole drug, posaconazole. The determination of the structures of trypanosomatid and human SQS enzymes with a diverse set of inhibitors active in cells provides insights into SQS inhibition, of interest in the context of the development of drugs against Chagas disease.
Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China.
Na
N
Shang
Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China; Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan.
Qian
Q
Li
Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China; Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan.
Tzu-Ping
TP
Ko
Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China.
Hsiu-Chien
HC
Chan
Center for Biophysics and Computational Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois, United States of America.
Jikun
J
Li
Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China.
Yingying
Y
Zheng
Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China.
Chun-Hsiang
CH
Huang
Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China.
Feifei
F
Ren
Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China.
Chun-Chi
CC
Chen
Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China.
Zhen
Z
Zhu
Center for Tropical and Emerging Global Diseases and Department of Cellular Biology, University of Georgia, Athens, Georgia, United States of America.
Melina
M
Galizzi
Center for Tropical and Emerging Global Diseases and Department of Cellular Biology, University of Georgia, Athens, Georgia, United States of America.
Zhu-Hong
ZH
Li
Instituto de Parasitología y Biomedicina "Lopez-Neyra", Consejo Superior de Investigaciones Cientificas, Granada, Spain.
Carlos A
CA
Rodrigues-Poveda
Instituto de Parasitología y Biomedicina "Lopez-Neyra", Consejo Superior de Investigaciones Cientificas, Granada, Spain.
Dolores
D
Gonzalez-Pacanowska
Laboratório de Ultraestrutura Celular Hertha Meyer, CCS, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Ilha do Fundão, Rio de Janeiro, Brazil; Instituto Nacional de Ciência e Tecnologia em Biologia Estrutural e Bioimagens, Universidade Federal do Rio de Janeiro, Cidade Universitária, Ilha do Fundão, Rio de Janeiro, Brazil; Diretoria de Programa, Instituto Nacional de Metrologia, Normalização e Qualidade Industrial-INMETRO, Duque de Caxias, Rio de Janeiro, Brazil.
Phercyles
P
Veiga-Santos
Laboratório de Ultraestrutura Celular Hertha Meyer, CCS, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Ilha do Fundão, Rio de Janeiro, Brazil; Instituto Nacional de Ciência e Tecnologia em Biologia Estrutural e Bioimagens, Universidade Federal do Rio de Janeiro, Cidade Universitária, Ilha do Fundão, Rio de Janeiro, Brazil; Diretoria de Programa, Instituto Nacional de Metrologia, Normalização e Qualidade Industrial-INMETRO, Duque de Caxias, Rio de Janeiro, Brazil.
Tecia Maria Ulisses
TM
de Carvalho
Laboratório de Ultraestrutura Celular Hertha Meyer, CCS, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Ilha do Fundão, Rio de Janeiro, Brazil; Instituto Nacional de Ciência e Tecnologia em Biologia Estrutural e Bioimagens, Universidade Federal do Rio de Janeiro, Cidade Universitária, Ilha do Fundão, Rio de Janeiro, Brazil; Diretoria de Programa, Instituto Nacional de Metrologia, Normalização e Qualidade Industrial-INMETRO, Duque de Caxias, Rio de Janeiro, Brazil.
Wanderley
W
de Souza
Instituto Venezolano de Investigaciones Cientificas, Caracas, Venezuela.
Julio A
JA
Urbina
Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan.
Andrew H-J
AH
Wang
Center for Tropical and Emerging Global Diseases and Department of Cellular Biology, University of Georgia, Athens, Georgia, United States of America.
Roberto
R
Docampo
Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois, United States of America.
Kai
K
Li
Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois, United States of America.
Yi-Liang
YL
Liu
Center for Biophysics and Computational Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois, United States of America; Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois, United States of America.
Eric
E
Oldfield
Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China.
Rey-Ting
RT
Guo
D000818:Animals; D014355:Chagas Disease / Q000188:drug therapy*; D002522:Chlorocebus aethiops; D018360:Crystallography, X-Ray; D004164:Diphosphonates / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology / Q000627:therapeutic use*; D013186:Farnesyl-Diphosphate Farnesyltransferase / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008958:Models, Molecular; D058990:Molecular Targeted Therapy / Q000379:methods*; D011106:Polyisoprenyl Phosphates / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011812:Quinuclidines / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D012717:Sesquiterpenes / Q000737:chemistry / Q000378:metabolism; D014344:Trypanocidal Agents / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology / Q000627:therapeutic use*; D014349:Trypanosoma cruzi / Q000201:enzymology; D014709:Vero Cells
D004164:Diphosphonates; D004791:Enzyme Inhibitors; D011106:Polyisoprenyl Phosphates; D011812:Quinuclidines; D012717:Sesquiterpenes; D014344:Trypanocidal Agents; C004808:farnesyl pyrophosphate; D013186:Farnesyl-Diphosphate Farnesyltransferase