Single-Molecule Force Spectroscopy Reveals that the Fe–N Bond Enables Multiple Rupture Pathways of the 2Fe2S Cluster in a MitoNEET Monomer

The mitochondrial outer membrane protein, mitoNEET (mNT), is an iron–sulfur protein containing an Fe2S2(His)1(Cys)3 cluster with a unique single Fe–N bond. Previous studies have shown that this Fe­(III)–N­(His) bond is essential for metal cluster transfer and protein function. To further understand...

Full description

Saved in:
Bibliographic Details
Published in:Analytical chemistry (Washington) 2020-11, Vol.92 (21), p.14783-14789
Main Authors: Song, Guobin, Ding, Xuan, Liu, Huaxing, Yuan, Guodong, Tian, Fang, Shi, Shengchao, Yang, Yang, Li, Guoqiang, Zheng, Peng
Format: Article
Language:English
Subjects:
Atomic force microscopy
Chemistry
Humans
Iron
Iron - chemistry
Iron-sulfur proteins
Membrane proteins
Metal clusters
Mitochondria
Mitochondrial Proteins - chemistry
Models, Molecular
Monomers
Nitrogen - chemistry
Protein Conformation
Proteins
Rupture
Rupturing
Single Molecule Imaging
Spectroscopy
Spectrum analysis
Sulfur
Sulfur - chemistry
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The mitochondrial outer membrane protein, mitoNEET (mNT), is an iron–sulfur protein containing an Fe2S2(His)1(Cys)3 cluster with a unique single Fe–N bond. Previous studies have shown that this Fe­(III)–N­(His) bond is essential for metal cluster transfer and protein function. To further understand the effect of this unique Fe–N bond on the metal cluster and protein, we used atomic force microscopy-based single-molecule force spectroscopy (AFM-SMFS) to investigate the mechanical unfolding mechanism of an mNT monomer, focusing on the rupture pathway and kinetic stability of the cluster. We found that the Fe–N bond was the weakest point of the cluster, the rupture of which occurred first, and could be independent of the cluster break. Moreover, this Fe–N bond enabled a dynamic and labile iron–sulfur cluster, as multiple unfolding pathways of mNT with a unique Fe2S2(Cys)3 intermediate were observed accordingly.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.0c03536