Biophysical Characterization of Iron-Sulfur Proteins

Bhanu P Jagilinki; Irina Paluy; Alexei M Tyryshkin; Vikas Nanda; Dror Noy

doi:10.21769/BioProtoc.4202

Biophysical Characterization of Iron-Sulfur Proteins

Bio Protoc. 2021 Oct 20;11(20):e4202. doi: 10.21769/BioProtoc.4202.

Authors

Bhanu P Jagilinki^{1

2}, Irina Paluy¹, Alexei M Tyryshkin², Vikas Nanda², Dror Noy¹

Affiliations

¹ Migal-Galilee Research Institute, Kiryat Shmona 11016, Israel.
² Center for Advanced Biotechnology and Medicine and the Department of Biochemistry and Molecular Biology, Robert Wood Johnson Medical School, Rutgers University, Piscataway, New Jersey 08854, United States.

Abstract

Iron-sulfur proteins are primordial catalysts and biological electron carriers that today drive major metabolic pathways across all forms of life. They can access a diversity of oxidation states and can mediate electron transfer over an extended range of reduction potentials spanning more than 1 V. Depending on the protein micro-environment and geometry of ligand, co-ordination the iron-sulfur clusters can occur in different forms [2Fe-2S], [3Fe-4S], HiPIP [4Fe-4S], and [4Fe-4S]. There are several spectroscopic methods available to characterize the composition and electronic configuration of the iron-sulfur clusters, such as optical methods and electron paramagnetic resonance. This paper presents the protocols used to characterize the metal center of Coiled-Coil Iron-Sulfur (CCIS), an artificial metalloprotein containing one [4Fe-4S] cluster. It is expected that these protocols will be of general utility for other iron-sulfur proteins.

Keywords: Electron Paramagnetic Resonance; Iron-sulfur proteins; UV-visible spectroscopy; [4Fe-4S] clusters; and ICP-AES.