Selection of synthetic proteins to modulate the human frataxin function. Issue 2 (28th October 2022)

• Record Type: Journal Article
• Authors: Pignataro, María Florencia; Herrera, María Georgina; Fernández, Natalia Brenda; Aran, Martín; Gentili, Hernán Gustavo; Battaglini, Fernando; Santos, Javier
• Is Part Of: Biotechnology and bioengineering. Volume 120:Issue 2(2023)
• Page Start: 409
• Page End: 425
• Publication Date: 2022-10-28
• Subjects: conformational stability -- iron‐sulfur cluster assembly -- protein–protein interaction; Biotechnology -- Periodicals; Bioengineering -- Periodicals; 660.6
• DOI: 10.1002/bit.28263
• Languages: English
• ISSNs: 0006-3592

Abstract

Abstract: Frataxin is a kinetic activator of the mitochondrial supercomplex for iron‐sulfur cluster assembly. Low frataxin expression or a decrease in its functionality results in Friedreich's Ataxia (FRDA). With the aim of creating new molecular tools to study this metabolic pathway, and ultimately, to explore new therapeutic strategies, we have investigated the possibility of obtaining small proteins exhibiting a high affinity for frataxin. In this study, we applied the ribosome display approach, using human frataxin as the target. We focused on Affi_224, one of the proteins that we were able to select after five rounds of selection. We have studied the interaction between both proteins and discussed some applications of this specific molecular tutor, concerning the modulation of the supercomplex activity. Affi_224 and frataxin showed a K D value in the nanomolar range, as judged by surface plasmon resonance analysis. Most likely, it binds to the frataxin acidic ridge, as suggested by the analysis of chemical shift perturbations (nuclear magnetic resonance) and computational simulations. Affi_224 was able to increase Cys NFS1 desulfurase activation exerted by the FRDA frataxin variant G130V. Importantly, Affi_224 interacts with frataxin in a human cellular model. Our results suggest quaternary addition may be a new tool to modulate frataxin function in vivo. Nevertheless, more functional experiments under physiological conditions should be carried out to evaluate Affi_224 effectiveness in FRDA cell models. Abstract : Frataxin is a kinetic activator for iron‐sulfur cluster assembly. Low frataxin expression or a decrease in its functionality results in Friedreich's Ataxia (FRDA). We applied the ribosome display approach, using human frataxin as the target to select affitins as tutor proteins. Affi_224 showed a nanomolar K D, it binds to the frataxin acidic ridge, and it was able to increase the activation exerted by the FRDA frataxin variant G130V. Moreover, Affi_224 interacts with frataxin in a cellular environment.