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

Karol Nass; Lars Redecke; M. Perbandt; O. Yefanov; M. Klinge; R. Koopmann; F. Stellato; A. Gabdulkhakov; R. Schönherr; D. Rehders; J. M. Lahey-Rudolph; A. Aquila; A. Barty; S. Basu; R. B. Doak; R. Duden; M. Frank; R. Fromme; S. Kassemeyer; G. Katona; R. Kirian; H. Liu; I. Majoul; J. M. Martin-Garcia; M. Messerschmidt; R. L. Shoeman; U. Weierstall; S. Westenhoff; T. A. White; G. J. Williams; C. H. Yoon; N. Zatsepin; P. Fromme; M. Duszenko; H. N. Chapman; C. Betzel

doi:10.1038/s41467-020-14484-w

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

Karol Nass, Lars Redecke, M. Perbandt, O. Yefanov, M. Klinge, R. Koopmann, F. Stellato, A. Gabdulkhakov, R. Schönherr, D. Rehders, J. M. Lahey-Rudolph, A. Aquila, A. Barty, S. Basu, R. B. Doak, R. Duden, M. Frank, R. Fromme, S. Kassemeyer, G. KatonaR. Kirian, H. Liu, I. Majoul, J. M. Martin-Garcia, M. Messerschmidt, R. L. Shoeman, U. Weierstall, S. Westenhoff, T. A. White, G. J. Williams, C. H. Yoon, N. Zatsepin, P. Fromme, M. Duszenko, H. N. Chapman, C. Betzel

Research output: Contribution to journal › Article › peer-review

24 Scopus citations

Abstract

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.

Original language	English (US)
Article number	620
Journal	Nature communications
Volume	11
Issue number	1
DOIs	https://doi.org/10.1038/s41467-020-14484-w
State	Published - Dec 1 2020

ASJC Scopus subject areas

General Chemistry
General Biochemistry, Genetics and Molecular Biology
General Physics and Astronomy

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Nass, K., Redecke, L., Perbandt, M., Yefanov, O., Klinge, M., Koopmann, R., Stellato, F., Gabdulkhakov, A., Schönherr, R., Rehders, D., Lahey-Rudolph, J. M., Aquila, A., Barty, A., Basu, S., Doak, R. B., Duden, R., Frank, M., Fromme, R., Kassemeyer, S., ... Betzel, C. (2020). In cellulo crystallization of Trypanosoma brucei IMP dehydrogenase enables the identification of genuine co-factors. Nature communications, 11(1), Article 620. https://doi.org/10.1038/s41467-020-14484-w

Nass, K, Redecke, L, Perbandt, M, Yefanov, O, Klinge, M, Koopmann, R, Stellato, F, Gabdulkhakov, A, Schönherr, R, Rehders, D, Lahey-Rudolph, JM, Aquila, A, Barty, A, Basu, S, Doak, RB, Duden, R, Frank, M, Fromme, R, Kassemeyer, S, Katona, G, Kirian, R, Liu, H, Majoul, I, Martin-Garcia, JM, Messerschmidt, M, Shoeman, RL, Weierstall, U, Westenhoff, S, White, TA, Williams, GJ, Yoon, CH, Zatsepin, N, Fromme, P, Duszenko, M, Chapman, HN & Betzel, C 2020, 'In cellulo crystallization of Trypanosoma brucei IMP dehydrogenase enables the identification of genuine co-factors', Nature communications, vol. 11, no. 1, 620. https://doi.org/10.1038/s41467-020-14484-w

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title = "In cellulo crystallization of Trypanosoma brucei IMP dehydrogenase enables the identification of genuine co-factors",

abstract = "Sleeping sickness is a fatal disease caused by the protozoan parasite Trypanosoma brucei (Tb). Inosine-5{\textquoteright}-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 {\AA} 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.",

author = "Karol Nass and Lars Redecke and M. Perbandt and O. Yefanov and M. Klinge and R. Koopmann and F. Stellato and A. Gabdulkhakov and R. Sch{\"o}nherr and D. Rehders and Lahey-Rudolph, {J. M.} and A. Aquila and A. Barty and S. Basu and Doak, {R. B.} and R. Duden and M. Frank and R. Fromme and S. Kassemeyer and G. Katona and R. Kirian and H. Liu and I. Majoul and Martin-Garcia, {J. M.} and M. Messerschmidt and Shoeman, {R. L.} and U. Weierstall and S. Westenhoff and White, {T. A.} and Williams, {G. J.} and Yoon, {C. H.} and N. Zatsepin and P. Fromme and M. Duszenko and Chapman, {H. N.} and C. Betzel",

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doi = "10.1038/s41467-020-14484-w",

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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.

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.

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SN - 2041-1723

VL - 11

JO - Nature communications

JF - Nature communications

IS - 1

M1 - 620

ER -

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

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In cellulo crystallization of Trypanosoma brucei IMP dehydrogenase enables the identification of ATP and GMP as genuine co-factors

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

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In cellulo crystallization of Trypanosoma brucei IMP dehydrogenase enables the identification of ATP and GMP as genuine co-factors

Cite this