Single‐Molecule Force Spectroscopy Reveals Stability of mitoNEET and its [2Fe2Se] Cluster in Weakly Acidic and Basic Solutions

• DOI: 10.1002/open.202200056
• Publication Date: 2022-05-24T10:03:17Z
• Authors: Jing‐Yuan Nie; Guo‐Bin Song; Yi‐Bing Deng; Peng Zheng
• Keywords: Force Spectroscopy; Chemical Stability

The outer mitochondrial membrane protein mitoNEET (mNT) is a recently identified iron-sulfur containing unique Fe2 S2 (His)1 (Cys)3 metal cluster with single Fe-N(His87) coordinating bond. This labile Fe-N bond led to multiple unfolding/rupture pathways of mNT and its by atomic force microscopy-based single-molecule spectroscopy (AFM-SMFS), one most common tools for characterizing the molecular mechanics. Although previous ensemble studies showed that this Fe-N(His) essential function, they also indicated [2Fe2S] are stable under acidic conditions. Thus, we applied AFM-SMFS measure stability at pH values 6, 7, 8. Indeed, all unfolding were still observed. Moreover, measurements revealed stabilities consistent these only ≈20 pN differences. found behavior in both weakly basic solutions despite