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In vitro PhoX cross-linking of purified hsMic60

ABSTRACT: Mitochondrial crista junctions (CJs) operate as regulated gateways into the cristae microenvironment, whose protein, metabolite, and ion compositions are finely tuned for mitochondrial function. The Mic60-Mic19 complex of the mitochondrial contact site and cristae organizing system (MICOS) complex was suggested to span across CJs and act as a diffusion barrier, but little is known of how its dynamic architecture facilitates this task. To address this open question, we determined the crystal structure of an amino-terminal dimeric helical bundle of human Mic60. These and previous structural and biochemical data were harnessed in molecular dynamic (MD) simulations to develop a dynamic model of the human tetrameric Mic60-Mic19 subcomplex in the CJ environment, to validate its architecture using in organello cross-linking data and to computationally characterize its function as a diffusion barrier. Our integrative structural biology approach enables the functional investigation of flexible, multidomain protein complexes which escape conventional structural biology methods. This in vitro PhoX cross-linking mass spectrometry data of purified hsMic60 supports the in organello studies by showing a similar cross-linking pattern.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Max Ruwolt

LAB HEAD: Oliver Daumke

PROVIDER: PXD077526 | Pride | 2026-08-04

REPOSITORIES: Pride

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			Action	DRS
	Ex1_20250331_MxR_MR_Evangelia_250325_M60_HS_18C_peak2_PhoX.xlsx	Xlsx
	Ex1_20250331_MxR_MR_Evangelia_250325_M60_HS_18C_peak2_PhoX_inj1.raw	Raw
	Ex1_20250331_MxR_MR_Evangelia_250325_M60_HS_18C_peak2_PhoX_inj2.raw	Raw
	MxR_Evangelia_M60HS18C.fasta	Fasta
	checksum.txt	Txt

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