Cellular-Structural Biology Lab


The structure and function of proteins depend on multiple factors such as sub-celullar localization, post-translational modifications and interactions with other biomolecules. However, most protein structural and mechanistic studies are typically performed on isolated samples, under conditions that differ substantially from in vivo environments of live cells. In our group we use NMR in live cells and multicellular organisms to obtain structural and functional information of proteins in their native environments and to study how oxidative post-translational modifications modulate central physiological and pathological processes involved in cardiovascular and neurodegenerative disorders.

Research Lines

Oxidative stress and heart development

A large number of congenital heart defects (CHD) arise as a consequence of the interaction between environmental factors, infections and genetic susceptibilities. Imbalances in oxidative regulation mechanisms result in cardiac pathologies. Exposure to oxidative stress during pregnancy has been linked with CHD and a detailed knowledge of its effects on cardiac development at the morphological, molecular and cellular level constitutes the basis for the development of early detection methods and the design of novel therapeutic strategies. In this research line, headed by Dr Verónica Lombardo, we use high resolution microscopy, molecular and cell biology and NMR methodologies in zebrafish (Danio rerio) embryos to understand how oxidative stress impacts on cardiac morphogenesis and elucidate the redox signaling pathways involved in the development of congenital heart defects.

Oxidative stress, lipids and oxidative protein modifications

The increase in reactive oxygen species and the decline of enzymatic redox protective systems have been identified as risk factors for the development of neurodegenerative and cardiovascular diseases. They elicit multiple effects including protein oxidative modifications interfering with normal function and changes in the lipid composition of cell membranes, which introduce cellular stress and metabolic imbalances that promote pathological states. In a collaborative effort between our group and the group of Dr. Diego de Mendoza, we analyze the role of oxidative stress and the metabolism of unsaturated fatty acids and cholesterol in the cellular aggregation of amyloid proteins and other pathological processes. We use NMR methodologies in vitro, e in vivo, in Caenorhabditis elegans and in zebrafish, combined with protein biochemistry, microscopy and metabolomics approaches. Our initiative seeks to obtain a comprehensive high resolution picture of the effect of oxidative stress on cellular pathologies.

Selected Publications

  • Megadalton-sized dityrosine aggregates of α-Synuclein retain high degrees of structural disorder and internal dynamics. Mol. Biol. 432, 166689, Verzini S., Shah M., Theillet F.X., Belsom A., Bieschke J., Wanker E.E., Rappsilber J., Binolfi A., Selenko P. (2020). https://doi.org/10.1016/j.jmb.2020.10.023
  • An NMR-based biosensor to measure stereo-specific methionine sulfoxide reductase (MSR) activities in vitro and in vivo. Eur. J. 26, 14838. Sánchez-López C., Labadie N.,  Lombardo V.A.,  Biglione F.A.,  Manta B., Jacob R., Gladyshev V., Abdelilah-Seyfried S., Selenko P., Binolfi A. (2020). https://doi.org/10.1002/chem.202002645
  • Morphogenetic control of zebrafish cardiac looping by Bmp signaling. Development, 146, 180091. Lombardo  A., Heise M., Moghtadaei M., Bornhorst D., Männer J., Abdelilah-Seyfried S. (2019). https://doi.org/10.1242/dev.180091 
  • Time-resolved NMR analysis of proteolytic α-synuclein processing in vitro and in cellulo. Proteomics, 18, 1800056. Limatola A., Eichmann C., Jacob R., Ben-Nissan G., Sharon M., Binolfi A., Selenko P. (2018). https://doi.org/10.1002/pmic.201800056
  • Structural disorder of monomeric α-synuclein persists in mammalian cells. Nature 530, 45. Theillet F-X., Binolfi A., Bekei B., Martorana A., Rose H. M., Stuiver M., Verzini S., Lorenz D., van Rossum M., Goldfarb D., Selenko P. (2016). https://doi.org/10.1038/nature16531
  • Intracellular repair of oxidation-damaged alpha-synuclein fails to target C-terminal modification sites. Commun. 7, 10251. Binolfi A., Limatola A., Verzini S., Kosten J., Theillet F-X., Rose H. M., Bekei B., Stuiver M., van Rossum M., Selenko P. (2016). https://doi.org/10.1038/ncomms10251
  • Copper binding to the N-terminally acetylated, naturally occurring form of alpha-synuclein induces local helical folding. Am. Chem. Soc. 137, 6444. Miotto M. C., Valiente-Gabioud A. A., Rossetti G., Zweckstetter M., Carloni P., Selenko P., Griesinger C., Binolfi A., Fernández C. O.* (2015). https://doi.org/10.1021/jacs.5b01911
  • Physicochemical and biological properties of cells and their effects on IDPs. Rev. 114, 6661. Theillet F-X., Binolfi A., Frembgen-Kesner T., Hingorani K., Sarkar M., Kyne C., Li C., Crowley P., Gierasch L., Pielak G., Elcock A. H., Gershenson A., Selenko P. (2014). https://doi.org/10.1021/cr400695p
  • Blood flow and Bmp signaling control endocardial chamber morphogenesis. Dev Cell.,30, 367. Dietrich A. C., Lombardo V. A., Veerkamp J., Priller F., Abdelilah-Seyfried S. (2014). https://doi.org/10.1016/j.devcel.2014.06.020 
  • Bacterial in-cell NMR of human a-synuclein: a disordered monomer by nature? Soc. Trans. 40, 950. Binolfi A., Theillet F-X., Selenko, P. (2012). https://doi.org/10.1042/BST20120096 

For a complete list, check my Google Scholar profile


Twitter: @ABinolfi