Ni-NTA Affinity Chromatography to Characterize Protein-Protein Interactions During Fe-S Cluster Biogenesis

Terrell D Carter; F Wayne Outten

doi:10.1007/978-1-0716-1605-5_7

Ni-NTA Affinity Chromatography to Characterize Protein-Protein Interactions During Fe-S Cluster Biogenesis

Methods Mol Biol. 2021:2353:125-136. doi: 10.1007/978-1-0716-1605-5_7.

Authors

Terrell D Carter¹, F Wayne Outten²

Affiliations

¹ Department of Chemistry and Biochemistry, University of South Carolina, Columbia, SC, USA.
² Department of Chemistry and Biochemistry, University of South Carolina, Columbia, SC, USA. woutten@sc.edu.

Abstract

Affinity chromatography can be repurposed to provide useful information about the specific partner protein(s) to which a protein of interest may bind as well as the relative binding affinity of that partner protein for the protein of interest. Here, we provide a protocol for an Ni-NTA affinity chromatography assay that may be utilized to uncover insightful information about the nature of protein-protein interactions during iron-sulfur (Fe-S) cluster biogenesis reactions.

Keywords: Affinity chromatography; Anaerobic; IMAC; Iron-sulfur cluster.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Chromatography, Affinity*
Iron / metabolism
Iron-Sulfur Proteins
Sulfur / metabolism

Substances

Iron-Sulfur Proteins
Sulfur
Iron

Abstract

Publication types

MeSH terms

Substances

Grants and funding