TY - JOUR
T1 - In cellulo crystallization of Trypanosoma brucei IMP dehydrogenase enables the identification of genuine co-factors
AU - Nass, Karol
AU - Redecke, Lars
AU - Perbandt, M.
AU - Yefanov, O.
AU - Klinge, M.
AU - Koopmann, R.
AU - Stellato, F.
AU - Gabdulkhakov, A.
AU - Schönherr, R.
AU - Rehders, D.
AU - Lahey-Rudolph, J. M.
AU - Aquila, A.
AU - Barty, A.
AU - Basu, S.
AU - Doak, R. B.
AU - Duden, R.
AU - Frank, M.
AU - Fromme, R.
AU - Kassemeyer, S.
AU - Katona, G.
AU - Kirian, R.
AU - Liu, H.
AU - Majoul, I.
AU - Martin-Garcia, J. M.
AU - Messerschmidt, M.
AU - Shoeman, R. L.
AU - Weierstall, U.
AU - Westenhoff, S.
AU - White, T. A.
AU - Williams, G. J.
AU - Yoon, C. H.
AU - Zatsepin, N.
AU - Fromme, P.
AU - Duszenko, M.
AU - Chapman, H. N.
AU - Betzel, C.
N1 - Funding Information:
XFEL experiments were carried out at the Linac Coherent Light Source (LCLS). LCLS is a national user facility operated by Stanford University on behalf of the U.S. Department of Energy, Office of Basic Energy Sciences. This work is in part supported by the Cluster of Excellence ‘The Hamburg Centre for Ultrafast Imaging’ of the Deutsche For-schungsgemeinschaft (DFG) – EXC 1074 – project ID 194651731, by the Helmholtz Excellence Network ‘Structure, Dynamics and Control on the Atomic Scale’, as well as by NSF award MCB-1021557. L.R., D.R., and C.B., and M.P. thank the German Federal Ministry for Education and Research (BMBF) for funding (grants 01KX0806, 01KX0807 and 05K16GUA) and the Joachim-Herz-Stiftung Hamburg via the project Infecto-Physics. L.R. thanks BMBF for funding via grant 05K18FLA. J.M.L. is grateful to the Joachim-Herz-Stiftung for a PhD scholarship. Support from the Hamburg Ministry of Science and Research and Joachim Herz Stiftung as part of the Hamburg Initiative for Excellence in Research (LEXI) and the Hamburg School for Structure and Dynamics (SDI) as well as the DFG Cluster of Excellence “Inflammation at Interfaces” (EXC 306) is gratefully acknowledged. Moreover, this research used resources of the National Synchrotron Light Source II, a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Brookhaven National Laboratory under Contract No. DE-SC0012704. This work was performed, in part, under the auspices of the US Department of Energy by Lawrence Livermore National Laboratory under the contract DE-AC52-07NA27344. M.F. was supported by the UCOP Lab Fee Program (award no. 118036) and LLNL LDRD project 12-ERD-031. G.K. was supported by the Swedish Research Council and the Knut and Alice Wallenberg Foundation. The authors thank Vadim Cherezov for acting as the spokesperson of the LCLS L490 beam time and Ilme Schlichting for helpful discussions.
Publisher Copyright:
© 2020, The Author(s).
Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 2020/12/1
Y1 - 2020/12/1
N2 - Sleeping 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.
AB - Sleeping 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.
UR - http://www.scopus.com/inward/record.url?scp=85078711387&partnerID=8YFLogxK
U2 - 10.1038/s41467-020-14484-w
DO - 10.1038/s41467-020-14484-w
M3 - Journal articles
C2 - 32001697
AN - SCOPUS:85078711387
SN - 1751-8628
VL - 11
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 620
ER -
