Biomolecular Fishing for Calixarene Partners by a Chemoproteomic Approach.
MS-based chemical-proteomics technology is introduced herein as a third general strategy to study the biomolecular recognition properties of given calixarene derivatives. In particular, we demonstrate that a simply designed calix[4]arene derivative 1 a bearing acetamido groups at the exo rim (pAC), when linked to a solid support, is able to fish out a specific protein (PDI protein) from a crude extract of HeLa cells. Western blot and surface plasmon resonance studies confirmed the direct interaction between PDI and the linker-free pAC derivative 1 b with considerable affinity, and in vitro tests showed its inhibition of PDI chaperone activity. In accordance with the role of PDI in a variety of human cancers, biological tests showed that pAC 1 b was cytotoxic and cytostatic toward CAL-27 and PC-3 cancer cell lines in vitro. Docking studies showed that H bonds and hydrophobic interactions contribute to the stabilization of the PDI/pAC complex.
Dipartimento di Chimica e Biologia "A. Zambelli", Università di Salerno, Via Giovanni Paolo II 132, 84084 Fisciano (Salerno, Italy).
Stefano
S
Tommasone
Dipartimento di Chimica e Biologia "A. Zambelli", Università di Salerno, Via Giovanni Paolo II 132, 84084 Fisciano (Salerno, Italy).
Carmen
C
Talotta
Dipartimento di Chimica e Biologia "A. Zambelli", Università di Salerno, Via Giovanni Paolo II 132, 84084 Fisciano (Salerno, Italy). cgaeta@unisa.it.
Carmine
C
Gaeta
Dipartimento di Farmacia, Università di Salerno, Via Giovanni Paolo II 132, 84084 Fisciano (Salerno, Italy).
Luigi
L
Margarucci
Dipartimento di Farmacia, Università di Salerno, Via Giovanni Paolo II 132, 84084 Fisciano (Salerno, Italy).
Maria Chiara
MC
Monti
Dipartimento di Farmacia, Università di Salerno, Via Giovanni Paolo II 132, 84084 Fisciano (Salerno, Italy). casapullo@unisa.it.
Agostino
A
Casapullo
Dipartimento di Medicina dei Sistemi, Università di Roma Tor Vergata, Via Montpellier 1, 00133 Roma (Italy).
Beatrice
B
Macchi
Dipartimento di Medicina dei Sistemi, Università di Roma Tor Vergata, Via Montpellier 1, 00133 Roma (Italy).
Salvatore Pasquale
SP
Prete
Department of Pathology, Laboratory of Dermatology and Immunodeficiencies, Medical School, University of Sao Paulo (Brasil).
Adriana
A
Ladeira De Araujo
Dipartimento di Chimica e Biologia "A. Zambelli", Università di Salerno, Via Giovanni Paolo II 132, 84084 Fisciano (Salerno, Italy). neri@unisa.it.
Placido
P
Neri
D047250:Calixarenes / Q000737:chemistry*; D003461:Crystallography; D011485:Protein Binding; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D040901:Proteomics
D011506:Proteins; D047250:Calixarenes
biomolecular recognition; calixarenes; chemical proteomics; proteins; surface plasmon resonance