A Combined NMR and Computational Approach to Determine the RGDechi-hCit-[alpha]v[beta]3 Integrin Recognition Mode in Isolated Cell Membranes

• Published in: Chemistry : a European journal Vol. 22; no. 2; p. 681
• Main Authors: Farina, Biancamaria, dePaola, Ivan, Russo, Luigi, Capasso, Domenica, Liguoro, Annamaria, Gatto, Annarita Del, Saviano, Michele, Pedone, Paolo V, DiGaetano, Sonia, Malgieri, Gaetano, Zaccaro, Laura, Fattorusso, Roberto
• Format: Journal Article
• Language: English
• Published: Weinheim Wiley Subscription Services, Inc 11-01-2016
• Subjects: Cell membranes; Chemistry; Computer applications; Design analysis; Determinants; Fragmentation; High resolution; In vitro methods and tests; Integrins; Internalization; Membranes; Metastases; NMR; Nuclear magnetic resonance; Pharmacology; Recognition
• ISSN: 0947-6539; 1521-3765
• DOI: 10.1002/chem.201503126

The critical role of integrins in tumor progression and metastasis has stimulated intense efforts to identify pharmacological agents that can modulate integrin function. In recent years, [alpha]v[beta]3 and [alpha]v[beta]5 integrin antagonists were demonstrated to be effective in blocking tumor progression. RGDechi-hCit, a chimeric peptide containing a cyclic RGD motif linked to an echistatin C-terminal fragment, is able to recognize selectively [alpha]v[beta]3 integrin both in vitro and in vivo. High-resolution molecular details of the selective [alpha]v[beta]3 recognition of the peptide are certainly required, nonetheless RGDechi-hCit internalization limited the use of classical in cell NMR experiments. To overcome such limitations, we used WM266 isolated cellular membranes to accomplish a detailed NMR interaction study that, combined with a computational analysis, provides significant structural insights into [alpha]v[beta]3 molecular recognition by RGDechi-hCit. Remarkably, on the basis of the identified molecular determinants, we design a RGDechi-hCit mutant that is selective for [alpha]v[beta]5 integrin.