Paramagnetic NMR to study iron sulfur proteins: ¹³C detected experiments illuminate the vicinity of the metal center
Creators
-
1.
University of Florence
-
2.
Universidade Nova de Lisboa
-
3.
California Institute of Technology
Abstract
The robustness of NMR coherence transfer in proximity of a paramagnetic center depends on the relaxation properties of the nuclei involved. In the case of Iron-Sulfur Proteins, different pulse schemes or different parameter sets often provide complementary results. Tailored versions of HCACO and CACO experiments significantly increase the number of observed C^α/C' connectivities in highly paramagnetic systems, by recovering many resonances that were lost due to paramagnetic relaxation. Optimized ¹³C direct detected experiments can significantly extend the available assignments, improving the overall knowledge of these systems. The different relaxation properties of C^α and C' nuclei are exploited in CACO vs COCA experiments and the complementarity of the two experiments is used to obtain structural information. The two [Fe₂S₂]⁺ clusters containing NEET protein CISD3 and the one [Fe₄S₄]²⁺ cluster containing HiPIP protein PioC have been taken as model systems. We show that tailored experiments contribute to decrease the blind sphere around the cluster, to extend resonance assignment of cluster bound cysteine residues and to retrieve details on the topology of the iron-bound ligand residues.
Copyright and License
© 2023 The Author(s). This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
Acknowledgement
The support of the CERM/CIRMMP center of Instruct-ERIC is gratefully acknowledged. This work was supported in part by the project "Potentiating the Italian Capacity for Structural Biology Services in Instruct-ERIC" (ITACA.SB, Project no. IR0000009) within the call MUR 3264/2021 PNRR M4/C2/L3.1.1, funded by the European Union—Next Generation EU. LQ is a PhD student under the Tuscany Health Ecosystem-ECS_00000017 (CUP B83C22003920001), spoke 7, funded by the European Union—Next Generation EU. This work was funded by national funds through FCT–Fundação para a Ciência e a Tecnologia, I.P. (FCT), Project MOSTMICRO-ITQB with refs UIDB/04612/2020 and UIDP/04612/2020, and LS4FUTURE Associated Laboratory (LA/P/0087/2020).
Funding
Open access funding provided by Università degli Studi di Firenze within the CRUI-CARE Agreement.
Contributions
MP, FC, LQ and ROL wrote the main manuscript text. LQ and JMS prepared figures. DG and IBT prepared the samples. LQ, DG, IBT and JMS collected and analyzed the data. All authors reviewed the manuscript.
Conflict of Interest
The authors declare no competing interests.
Files
10858_2023_Article_425.pdf
Files
(2.2 MB)
| Name | Size | Download all |
|---|---|---|
|
md5:cdb0e8af84beef054411a8d56137f933
|
33.5 kB | Download |
|
md5:87006e022bc24ba457130aaa7ffeb953
|
2.1 MB | Preview Download |
Additional details
Identifiers
- ISSN
- 1573-5001
- DOI
- 10.1007/s10858-023-00425-4
- PMCID
- PMC10687126
Related works
Funding
- Centro di Risonanze Magnetiche
- European Research Council