{"id":2290,"date":"2026-05-08T16:20:56","date_gmt":"2026-05-08T14:20:56","guid":{"rendered":"https:\/\/sites.unica.it\/pe2026\/?p=2290"},"modified":"2026-05-08T17:58:28","modified_gmt":"2026-05-08T15:58:28","slug":"valeria-la-gatta","status":"publish","type":"post","link":"https:\/\/sites.unica.it\/pe2026\/2026\/05\/08\/valeria-la-gatta\/","title":{"rendered":"Valeria La Gatta"},"content":{"rendered":"\n
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PC19 – Valeria La Gatta<\/strong><\/p>\n\n\n\n

University of Modena and Reggio Emilia, Italy<\/p>\n\n\n\n

LinkedIn<\/a><\/strong><\/p>\n\n\n\n

valeria.lagatta@unimore.it<\/strong><\/p>\n<\/div><\/div>\n\n\n\n

Characterization of human thymidylate synthase dimer disruptor interactions by photoaffinity labeling<\/strong><\/strong><\/td><\/tr>
La Gatta Valeria<\/u>1<\/sup>, <\/strong>Tagliazucchi Lorenzo2<\/sup>, Aiello Daniele1<\/sup>, Illuminati Davide3<\/sup>, Pacifico Salvatore3<\/sup>, Guerrini Remo3<\/sup>, and Costi Maria Paola1<\/sup><\/em>

1<\/sup><\/em> Department of Life Science, University of Modena and Reggio Emilia;
2<\/em><\/sup> Department of biotechnology, chemistry and pharmacy, University of Parma;
3<\/em> <\/sup>University of Ferrara.<\/td><\/tr>
Abstract<\/strong>
Human thymidylate synthase (hTS) is an essential enzyme in de novo DNA biosynthesis and a validated anticancer target. hTS exists in a dynamic equilibrium between an active dimer and an inactive monomer involved in autoregulatory control of protein expression [1]. Conventional active-site inhibitors induce hTS overexpression and drug resistance [1,2]. To overcome these limitations, we developed small molecules targeting the hTS dimer interface, acting as dimer destabilizers that inhibit enzymatic activity while preserving regulatory functions [2]. To investigate ligand-protein interactions and assess selectivity, a photoaffinity labeling\u2013mass spectrometry (PAL-MS) strategy was established. PAL employs photoreactive probes containing diazirine groups that, upon UV irradiation, generate reactive intermediates able to covalently trap proximal residues. This approach enables investigation of non-covalent and transient interactions, particularly suitable for interface-targeting ligands. The incorporation of an alkyne handle further allows bioorthogonal conjugation with reporter tags via copper-catalyzed azide\u2013alkyne cycloaddition, enabling enrichment and identification of labeled proteins by LC\u2013MS [3]. Among the interface-targeting compounds previously developed in our laboratory, A40 was selected as a scaffold for probe design due to its favorable solubility and synthetic accessibility. A diazirine-containing analogue was synthesized and successfully applied in PAL experiments on recombinant hTS, enabling mapping of the binding region at the dimer interface. To extend it to a cellular context, alkyne-functionalized derivatives were designed. A series of pre-probe analogues was prepared to mimic the steric and electronic properties of the final constructs. Among these, A40_3pre retained the highest inhibitory activity and was selected as precursor for the development of a fully functionalized probe suitable for cellular PAL-MS studies. This work establishes an integrated medicinal chemistry and chemical biology platform for the investigation of hTS dimer destabilizers. By combining rational probe design with PAL-MS, this approach enables characterization of ligand\u2013protein interactions at the dimer interface and supports proteome-wide selectivity assessment, providing a foundation for next-generation anticancer agents targeting thymidylate synthase.<\/td><\/tr>
References \u00a0<\/strong>
[1] Taddia, L.; D\u2019Arca, D.; Ferrari, S.; et al. Inside the biochemical pathways of thymidylate synthase perturbed by anticancer drugs: Novel strategies to overcome cancer chemoresistance.\u00a0Drug Resist. Updat.<\/em>\u00a02015<\/strong>, 23, 20\u201354. DOI: 10.1016\/j.drup.2015.10.003.
[2] Costantino, L.; Ferrari, S.; Santucci, M.; et al. Destabilizers of the thymidylate synthase homodimer accelerate its proteasomal degradation and inhibit cancer growth.\u00a0eLife<\/em>\u00a02022<\/strong>, 11, e73862. DOI: 10.7554\/eLife.73862.
[3] Smith, E.; Collins, I. Photoaffinity labeling in target- and binding-site identification.\u00a0Future Med. Chem.<\/em>\u00a02015<\/strong>, 7, 159\u2013183. DOI: 10.4155\/fmc.14.152.<\/td><\/tr><\/tbody><\/table><\/div><\/figure>\n\n\n\n

<\/p>\n","protected":false},"excerpt":{"rendered":"

PC19 – Valeria La Gatta University of Modena and Reggio Emilia, Italy LinkedIn valeria.lagatta@unimore.it Characterization of human thymidylate synthase dimer disruptor interactions by photoaffinity labeling La Gatta Valeria1, Tagliazucchi Lorenzo2, Aiello Daniele1, Illuminati Davide3, Pacifico Salvatore3, […]<\/p>\n","protected":false},"author":10371,"featured_media":0,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[1],"tags":[],"class_list":["post-2290","post","type-post","status-publish","format-standard","hentry","category-senza-categoria"],"_links":{"self":[{"href":"https:\/\/sites.unica.it\/pe2026\/wp-json\/wp\/v2\/posts\/2290","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/sites.unica.it\/pe2026\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/sites.unica.it\/pe2026\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/sites.unica.it\/pe2026\/wp-json\/wp\/v2\/users\/10371"}],"replies":[{"embeddable":true,"href":"https:\/\/sites.unica.it\/pe2026\/wp-json\/wp\/v2\/comments?post=2290"}],"version-history":[{"count":4,"href":"https:\/\/sites.unica.it\/pe2026\/wp-json\/wp\/v2\/posts\/2290\/revisions"}],"predecessor-version":[{"id":2368,"href":"https:\/\/sites.unica.it\/pe2026\/wp-json\/wp\/v2\/posts\/2290\/revisions\/2368"}],"wp:attachment":[{"href":"https:\/\/sites.unica.it\/pe2026\/wp-json\/wp\/v2\/media?parent=2290"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/sites.unica.it\/pe2026\/wp-json\/wp\/v2\/categories?post=2290"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/sites.unica.it\/pe2026\/wp-json\/wp\/v2\/tags?post=2290"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}