Studying GGDEF Domain in the Act: Minimize Conformational Frustration to Prevent Artefacts

• Published in: Life (Basel, Switzerland) Vol. 11; no. 1; p. 31
• Main Authors: Mantoni, Federico, Scribani Rossi, Chiara, Paiardini, Alessandro, Di Matteo, Adele, Cappellacci, Loredana, Petrelli, Riccardo, Ricciutelli, Massimo, Paone, Alessio, Cutruzzolà, Francesca, Giardina, Giorgio, Rinaldo, Serena
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 06.01.2021; MDPI
• Subjects: Allosteric properties; allostery; Biofilms; c-di-GMP; Catalysis; Dimerization; Domains; enzyimatic assay; Enzymes; GGDEF; Mass spectrometry; protein engineering; Protein expression; Proteins; Regulation; Scientific imaging; Signal transduction; United States > US; Germany; Chicago Illinois; enzyimatic assay; c-di-GMP; protein engineering; GGDEF; allostery
• ISSN: 2075-1729; 2075-1729
• DOI: 10.3390/life11010031

GGDEF-containing proteins respond to different environmental cues to finely modulate cyclic diguanylate (c-di-GMP) levels in time and space, making the allosteric control a distinctive trait of the corresponding proteins. The diguanylate cyclase mechanism is emblematic of this control: two GGDEF domains, each binding one GTP molecule, must dimerize to enter catalysis and yield c-di-GMP. The need for dimerization makes the GGDEF domain an ideal conformational switch in multidomain proteins. A re-evaluation of the kinetic profile of previously characterized GGDEF domains indicated that they are also able to convert GTP to GMP: this unexpected reactivity occurs when conformational issues hamper the cyclase activity. These results create new questions regarding the characterization and engineering of these proteins for in solution or structural studies.