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Title: In cellulo crystallization of Trypanosoma brucei IMP dehydrogenase enables the identification of genuine co-factors

Journal Article · · Nature Communications
ORCiD logo [1];  [2]; ORCiD logo [3];  [4];  [5]; ORCiD logo [6];  [7];  [8];  [9];  [10]; ORCiD logo [11];  [12];  [4];  [13];  [14];  [15]; ORCiD logo [16]; ORCiD logo [17];  [18]; ORCiD logo [19] more »; ORCiD logo [17]; ORCiD logo [20];  [15]; ORCiD logo [17];  [21]; ORCiD logo [18];  [17];  [19]; ORCiD logo [4];  [22];  [12]; ORCiD logo [23];  [17];  [6]; ORCiD logo [24];  [3] « less
  1. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Paul Scherrer Inst. (PSI), Villigen (Switzerland)
  2. Inst. of Biochemistry and Molecular Biology, Hamburg (Germany). Joint Lab. for Structural Biology of Infection and Inflammation; Univ. of Hamburg (Germany); Univ. of Lübeck, at Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Univ. of Lübeck (Germany); Deutsches Elektronen Synchrotron (DESY), Hamburg (Germany)
  3. Univ. of Hamburg, at Deutsches Elektronen-Synchrotron (DESY) (Germany); The Hamburg Centre for Ultrafast Imaging (CUI), Hamburg (Germany)
  4. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany)
  5. Inst. of Biochemistry and Molecular Biology, Hamburg (Germany). Joint Lab. for Structural Biology of Infection and Inflammation; Univ. of Hamburg (Germany); Univ. of Lübeck, at Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); BioAgilytix Europe GmbH, Hamburg (Germany)
  6. Univ. of Tübingen (Germany)
  7. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Univ. di Roma Tor Vergata, Rome (Italy); Istituto Nazionale di Fisica Nucleare (INFN), Rome (Italy)
  8. Russian Academy of Sciences, Moscow Region (Russia)
  9. Univ. of Lübeck (Germany); Deutsches Elektronen Synchrotron (DESY), Hamburg (Germany)
  10. Inst. of Biochemistry and Molecular Biology, Hamburg (Germany). Joint Lab. for Structural Biology of Infection and Inflammation; Univ. of Hamburg (Germany); Univ. of Lübeck, at Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); BODE Chemie GmbH, Hamburg (Germany)
  11. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Univ. of Lübeck (Germany)
  12. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); SLAC National Accelerator Lab., Menlo Park, CA (United States). Linac Coherent Light Source (LCLS)
  13. Arizona State Univ., Tempe, AZ (United States); European Molecular Biology Lab. (EMBL), Grenoble (France)
  14. Arizona State Univ., Tempe, AZ (United States); Max Planck Inst. for Medical Research, Heidelberg (Germany)
  15. Univ. of Lübeck (Germany)
  16. Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
  17. Arizona State Univ., Tempe, AZ (United States)
  18. Max-Planck-Inst. for Medical Research, Heidelberg (Germany)
  19. Univ. of Gothenburg (Sweden)
  20. Arizona State Univ., Tempe, AZ (United States); Beijing Computational Science Research Center (China)
  21. SLAC National Accelerator Lab., Menlo Park, CA (United States). Linac Coherent Light Source (LCLS); Arizona State Univ., Tempe, AZ (United States)
  22. SLAC National Accelerator Lab., Menlo Park, CA (United States). Linac Coherent Light Source (LCLS); Brookhaven National Lab. (BNL), Upton, NY (United States)
  23. Arizona State Univ., Tempe, AZ (United States); La Trobe Univ., VIC (Australia)
  24. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); The Hamburg Centre for Ultrafast Imaging (CUI), Hamburg (Germany); Univ. of Hamburg (Germany)
+ Show Author Affiliations

AbstractSleeping sickness is a fatal disease caused by the protozoan parasite Trypanosoma brucei (Tb). Inosine-5’-monophosphate dehydrogenase (IMPDH) has been proposed as a potential drug target, since it maintains the balance between guanylate deoxynucleotide and ribonucleotide levels that is pivotal for the parasite. Here we report the structure of TbIMPDH at room temperature utilizing free-electron laser radiation on crystals grown in living insect cells. The 2.80 Å resolution structure reveals the presence of ATP and GMP at the canonical sites of the Bateman domains, the latter in a so far unknown coordination mode. Consistent with previously reported IMPDH complexes harboring guanosine nucleotides at the second canonical site, TbIMPDH forms a compact oligomer structure, supporting a nucleotide-controlled conformational switch that allosterically modulates the catalytic activity. The oligomeric TbIMPDH structure we present here reveals the potential of in cellulo crystallization to identify genuine allosteric co-factors from a natural reservoir of specific compounds.

Research Organization:
Brookhaven National Laboratory (BNL), Upton, NY (United States). National Synchrotron Light Source II (NSLS-II); Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States); SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States). Linac Coherent Light Source (LCLS)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES); German Research Foundation (DFG); Helmholtz Excellence Network; National Science Foundation (NSF); German Federal Ministry for Education and Research (BMBF); Joachim-Herz-Stiftung Hamburg; UCOP Lab Fee Program; USDOE Laboratory Directed Research and Development (LDRD) Program; Swedish Research Council (VR); Knut and Alice Wallenberg Foundation; USDOE National Nuclear Security Administration (NNSA)
Grant/Contract Number:
SC0012704; MCB-1021557; 01KX0806; 01KX0807; 05K16GUA; 05K18FLA; AC52-07NA27344; 118036; AC02-76SF00515
OSTI ID:
1603290
Alternate ID(s):
OSTI ID: 1605174; OSTI ID: 2007604
Report Number(s):
BNL-213686-2020-JAAM; LLNL-JRNL-855421
Journal Information:
Nature Communications, Vol. 11, Issue 1; ISSN 2041-1723
Publisher:
Nature Publishing GroupCopyright Statement
Country of Publication:
United States
Language:
English

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