Modulation of the Structural and Functional Dynamics of the human mitochondrial protein ISCU2 using Nanoantibodies

NAIRA RODRIGUEZ; JULIÁN GROSSI; M. FLORENCIA PIGNATARO; MARÍA FLORENCIA PAVAN; LORENA ARCE; DANIELA MILITELLO; ITATÍ IBAÑEZ; JAVIER SANTOS

Córdoba

Congreso; Reunion Anual de la Sociedad Argentina de Biofísica; 2023

Sociedad Argentina de Biofísica

Background and objective: ISCU2-deficient myopathy is an inherited disease that primarilyaffects skeletal muscles. This myopathy is due to mutations in a gene that encodes theISCU2 protein, involved in the formation of FeS centers, is critical for the function ofmany different proteins, including those necessary for DNA repair and cellular respiration. Inthis work, we preliminarily selected and studied three different anti-ISCU2 nanoantibodies(NBs). By studying the interaction of ISCU2 with these NBs, we sought to modulate theactivity of the protein and the stabilization in its biologically active form.Methods: Nanobody libraries were prepared and several nanobodies showing an affinity forISCU2 were selected using the phage display technology. The interaction was evaluated bySEC-FLPC, ELISA and bioinformatics methods.Results: The predictions of the interaction of two NBs under study showed a certain sterichindrance, while the prediction of a NB (2A9) showed a more favorable union. It was thendecided to study the interaction of NB 2A9 with ISCU2 in vitro. This interaction doesn´t seemto affect the activity of ISCU2.Conclusions: We are investigating whether this NB could successfully stabilize unstableISCU2 variants in the context of the supercomplex. Furthermore, we think that these newtools will contribute to the understanding of the role that ISCU2 plays in Fe-S clusterbiosynthesis in the cellular environment.