Czech National Centre of the European Research Infrastructure Consortium INSTRUCT ERIC

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A gateway to realm of structural data for biochemists, biophysicists, molecular biologist, and all scientists whose research benefits from accurate structure determination of biological macromolecules, assemblies, and complex molecular machineries at atomic resolution.

Open access to 10 high-end core facilities and assisted expertise in NMR, X-ray crystallography and crystallization, cryo-electron microscopy and tomography, biophysical characterization of biomolecular interaction, nanobiotechnology, proteomics and structural mass spectrometry.

A distributed infrastructure constituted by Core Facilities of CEITEC (Central European Institute of Technology), located in Brno, and BIOCEV (Biotechnology and Biomedicine Centre), located in Vestec near Prague, Central Bohemia.

CIISB Core Facilities Research Highlights 

A. Schenkmayerova, et al.: Catalytic mechanism for Renilla-type luciferases, Nat. Catal. (2023). https://doi.org/10.1038/s41929-022-00895-z

M. Stiborek, et al.: Infrared Laser Desorption of Intact Nanoparticles for Digital Tissue Imaging Anal. Chem. (2022) 94, 51, 18114 – 18121, https://doi.org/10.1021/acs.analchem.2c05216

P.M. Szczepanik, et al.: Convergent Assembly of the Tricyclic Labdane Core Enables Synthesis of Diverse Forskolin-like Molecules, Angew. Chem. Int. Ed. (2022), https://doi.org/10.1002/anie.202213183

V. Siahaan, et al.: Microtubule lattice spacing governs cohesive envelope formation of tau family proteins, Nat. Chem. Biol., 18 (2022) 1224-+, 10.1038/s41589-022-01096-2

O. Gahura, et al.: An ancestral interaction module promotes oligomerization in divergent mitochondrial ATP synthases, Nature Communications, 13 (2022) 13, 10.1038/s41467-022-33588-z

More publications

CIISB Research Highlights

the best of science obtained using CIISB Core Facilities

  • Nature Catalysis 2023

    Nature Catalysis 2023

    The mechanism of Renilla-type luciferase reaction and its inhibition.

    Martin Marek and Zbyněk Prokop Research Group

    Significance

    The widely used coelenterazine-powered Renilla luciferase was discovered over 40 years ago, but the oxidative mechanism by which it generates blue photons remains unclear. Here we decipher Renilla-type catalysis through crystallographic, spectroscopic and computational experiments. Structures of ancestral and extant luciferases complexed with the substrate-like analogue azacoelenterazine or a reaction product were obtained, providing molecular snapshots of coelenterazine-to-coelenteramide oxidation. Bound coelenterazine adopts a Y-shaped conformation, enabling the deprotonated imidazopyrazinone component to attack O2 via a radical charge-transfer mechanism. A high emission intensity is secured by an aspartate from a conserved proton-relay system, which protonates the excited coelenteramide product. Another aspartate on the rim of the catalytic pocket fine-tunes the electronic state of coelenteramide and promotes the formation of the blue light-emitting phenolate anion. The results obtained also reveal structural features distinguishing flash-type from glow-type bioluminescence, providing insights that will guide the engineering of next-generation luciferase‒luciferin pairs for ultrasensitive optical bioassays.

     

    Schenkmayerova, A., Toul, M., Pluskal, D., Baatallah, R., Gagnot, G., Pinto, G.P., Santana, V.T., Stuchla, M., Neugebauer, P., Chaiyen, P., Damborsky, J., Bednar, D., Janin, Y.L., Prokop, Z. & Marek, M.: Catalytic mechanism for Renilla-type luciferases, Nat. Catal. (2023). https://doi.org/10.1038/s41929-022-00895-z

  • Analytical Chemistry 2022

    Analytical Chemistry 2022

    A new technique for the digital mapping of biomarkers in tissues based on desorption and counting intact gold nanoparticle (Au NP) tags using infrared laser ablation single - particle inductively coupled plasma mass spectrometry (IR LA SP ICP MS).

    Jan Preisler Research Group

    Significance

    In contrast to conventional UV laser ablation, Au NPs are not disintegrated during the desorption process due to their low absorption at 2940 nm. A mass spectrometer detects up to 83% of Au NPs. The technique is demonstrated on mapping a proliferation marker, nuclear protein Ki-67, in three-dimensional (3D) aggregates of colorectal carcinoma cells, and the results are compared with confocal fluorescence microscopy and UV LA ICP MS. Precise counting of 20 nm Au NPs with a single-particle detection limit in each pixel by the new approach generates sharp distribution maps of a specific biomarker in the tissue. Advantageously, the desorption of Au NPs from regions outside the tissue is strongly suppressed. The developed methodology promises multiplex mapping of low-abundant biomarkers in numerous biological and medical applications using multielemental mass spectrometers.

    Stiborek, M., Jindřichová, L., Meliorisová, S., Bednařík, A., Prysiazhnyi, V., Kroupa, J., Houška, P., Adamová, B., Navrátilová, J., Kanický, V., and Preisler, J.:

    Infrared Laser Desorption of Intact Nanoparticles for Digital Tissue Imaging

    Anal. Chem. 2022 94, 51, 18114 – 18121, https://doi.org/10.1021/acs.analchem.2c05216

  • Angewandte Chemie Int. Ed. 2022

    Angewandte Chemie Int. Ed. 2022

    Labdane-type terpenes are a large family of bioactive natural products. Their often-complex structures present challenges to semisynthesis and de novo chemical synthesis in search of analogs with improved properties. To enable new modifications of the well-known tricyclic terpene forskolin, we have developed a distinct and potentially general synthetic scheme for the preparation of analogs of complex labdanes.

    Jakub Švenda Research Group

    Significance

    We report a new synthetic strategy for the flexible preparation of forskolin-like molecules. The approach is different from the previously published works and employs a convergent assembly of the tricyclic labdane-type core from pre-functionalized cyclic building blocks. Stereoselective Michael addition enabled the fragment coupling with excellent control over three newly created contiguous stereocenters, all-carbon quaternary centers included. Silyl enol ether-promoted ring-opening metathesis paired with ring closure were the other key steps enabling concise assembly of the tricyclic core. Late-stage functionalization sequences transformed the tricyclic intermediates into a set of different forskolin-like molecules. The modular nature of the synthetic scheme described herein has the potential to become a general platform for the preparation of analogs of forskolin and other complex tricyclic labdanes.

    Szczepanik, P. M., Mikhaylov, A. A., Hylse, O., Kučera, R., Daďová, P., Nečas, M., Kubala, L., Paruch, K., and Švenda, J.:

    Convergent Assembly of the Tricyclic Labdane Core Enables Synthesis of Diverse Forskolin-like Molecules, Angew. Chem. Int. Ed. 2022, on-line version e202213183, https://doi.org/10.1002/anie.202213183

More publications Research Highlights archive

Reader’s Corner

literature to read, science to follow

In this section, a distinct selection of six highly stimulating research publications and reviews published during past 6 months is presented. It is our hope that links to exciting science, which deserves attention of the structural biology community, will help you to locate gems in the steadily expanding jungle of scientific literature. You are welcome to point out to any paper you found interesting by sending a link or citation to readerscorner@ciisb.org. The section is being updated regularly.


 

Reader’s Corner Archive

Quote of January

“Science is a wonderful thing if one does not have to earn one's living at it.”

Albert Einstein

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