biostudies-literatureSyed ANIAID NIH HHSNational Natural Science Foundation of ChinaNIHNIH Common FundNIGMS NIH HHS575-584.e3https://www.ebi.ac.uk/biostudies/studies/S-EPMC11069440biostudies-literatureUnknown32(5)Chaperonins Hsp60s are required for cellular vitality by assisting protein folding in an ATP-dependent mechanism. Although conserved, the human mitochondrial mHsp60 exhibits molecular characteristics distinct from the E. coli GroEL, with different conformational assembly and higher subunit association dynamics, suggesting a different mechanism. We previously found that the pathological mutant mHsp60^V72I exhibits enhanced subunit association stability and ATPase activity. To provide structural explanations for the V72I mutational effects, here we determined a cryo-EM structure of mHsp60^V72I. Our structural analysis combined with molecular dynamic simulations showed mHsp60^V72I with increased inter-subunit interface, binding free energy, and dissociation forceStructure (London, England : 1993)Cryo-EM structure and molecular dynamic simulations explain the enhanced stability and ATP activity of the pathological chaperonin mutant.PMC11069440U24 GM129539R01 AI173104Wang JCChen LZhai JSyed AGuo BZhao YfalseCryo-EM structure and molecular dynamic simulations explain the enhanced stability and ATP activity of the pathological chaperonin mutant.Chaperonins Hsp60s are required for cellular vitality by assisting protein folding in an ATP-dependent mechanism. Although conserved, the human mitochondrial mHsp60 exhibits molecular characteristics distinct from the E. coli GroEL, with different conformational assembly and higher subunit association dynamics, suggesting a different mechanism. We previously found that the pathological mutant mHsp60^V72I exhibits enhanced subunit association stability and ATPase activity. To provide structural explanations for the V72I mutational effects, here we determined a cryo-EM structure of mHsp60^V72I. Our structural analysis combined with molecular dynamic simulations showed mHsp60^V72I with increased inter-subunit interface, binding free energy, and dissociation force2024-01-01T00:00:00Z2024 May2026-06-02T20:56:11.511Z2025-07-06T03:05:51.298ZS-EPMC110694403841285510.1016/j.str.2024.02.001