The Mitochondrial Intermembrane Space Oxireductase Mia40 Funnels the Oxidative Folding Pathway of the Cytochrome c Oxidase Assembly Protein Cox19

Fraga, Hugo; Bech-Serra, Joan Josep; Canals, Françesc; Ortega, Gabriel; Millet, Óscar; Ventura, Salvador

doi:10.1074/jbc.m114.553479

Cited by 16 publications

(16 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In this system, the CSD does not discriminates between the 1‐SS and the 2‐SS forms, which can be separated, instead, by reversed‐phase chromatography . This is in line with previous evidence deriving from protein chemical modification and mutant analysis …”

Section: Comparison With Solution Methodssupporting

confidence: 87%

“…[56] This is in line with previous evidence deriving from protein chemical modification and mutant analysis. [57] The nascent polypeptide-associated complex (NAC) is a ribosome-associated chaperone important for protein homeostasis. Its native-MS spectrum displays a broad and multimodal CSD, dominated by peaks of the αβ heterodimer.…”

Section: Comparison With Solution Methodsmentioning

confidence: 99%

“…The line represents the linear fitting of the experimental results. B) SASA versus molecular weight relationship for globular proteins (blue curve, A s = 4.84 × M 0.760 ) and fully disordered proteins (red curve, A s = 1.29 × M 0.978 ) . The terms involved in CI definition are represented for a generic point on the plot.…”

Section: Compaction Indexmentioning

confidence: 99%

See 2 more Smart Citations

Conformational Characterization and Classification of Intrinsically Disordered Proteins by Native Mass Spectrometry and Charge‐State Distribution Analysis

et al. 2018

View full text Add to dashboard Cite

Intrinsically disordered proteins (IDPs) are systematically under‐represented in structural proteomics studies. Their structural characterization implies description of the dynamic conformational ensembles populated by these polymers in solution, posing major challenges to biophysical methods. “Native” MS (native‐MS) has emerged as a central tool in this field, conjugating the unique MS analytical power with structurally meaningful descriptors, like solvent‐accessible surface area (SASA) and collisional cross section (CCS). This review summarizes recently published papers comparing native‐MS and solution methods, with a focus on charge‐state‐distribution (CSD) analysis for IDP conformational analysis. The results point to substantial agreement, supporting structural interpretation of native‐MS spectra of IDPs. The discussion is integrated with data from our group on “synthetic” IDPs, obtained by reduction and alkylation of natively folded proteins, whose fold is stabilized by disulfide bridges. Finally, an MS‐based compaction index (CI) is introduced, evaluating SASA with reference to globular and fully disorder proteins. Such a parameter can be calculated for single conformers or the whole conformational ensemble, offering a continuous index for IDP comparison and classification.

show abstract

Section: Comparison With Solution Methodssupporting

confidence: 87%

Section: Comparison With Solution Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Conformational Characterization and Classification of Intrinsically Disordered Proteins by Native Mass Spectrometry and Charge‐State Distribution Analysis

et al. 2018

View full text Add to dashboard Cite

show abstract

“…First, a biologically relevant unfolded state can be accessed, since in their reduced state these proteins remain essentially unstructured, both in vitro and inside cells 13 . Second, non-catalyzed folding of these proteins can take several hours to occur 14 which permits tracking the process in real-time by a battery of low temporal-resolution techniques reporting on different structural properties. Third, in these proteins the adoption of the native structure and the formation of the disulfide bonds are linked in a process known as oxidative folding, allowing an accurate characterization of the major transient intermediates that populate the folding reaction 15 .…”

Section: Introductionmentioning

confidence: 99%

Disulfide driven folding for a conditionally disordered protein

Fraga

Pujols

Gil-García

et al. 2017

Sci Rep

Self Cite

View full text Add to dashboard Cite

Conditionally disordered proteins are either ordered or disordered depending on the environmental context. The substrates of the mitochondrial intermembrane space (IMS) oxidoreductase Mia40 are synthesized on cytosolic ribosomes and diffuse as intrinsically disordered proteins to the IMS, where they fold into their functional conformations; behaving thus as conditionally disordered proteins. It is not clear how the sequences of these polypeptides encode at the same time for their ability to adopt a folded structure and to remain unfolded. Here we characterize the disorder-to-order transition of a Mia40 substrate, the human small copper chaperone Cox17. Using an integrated real-time approach, including chromatography, fluorescence, CD, FTIR, SAXS, NMR, and MS analysis, we demonstrate that in this mitochondrial protein, the conformational switch between disordered and folded states is controlled by the formation of a single disulfide bond, both in the presence and in the absence of Mia40. We provide molecular details on how the folding of a conditionally disordered protein is tightly regulated in time and space, in such a way that the same sequence is competent for protein translocation and activity.

show abstract

“…Non-native disulfides can act as kinetic traps in protein folding and misfolding (30)(31)(32)(33). Hence, some pathways of protein aggregation depend strongly on the redox environment.…”

mentioning

confidence: 99%

An Internal Disulfide Locks a Misfolded Aggregation-Prone Intermediate in Cataract-Linked Mutants of Human Gamma-D Crystallin

Serebryany

Woodard

Adkar

et al. 2017

Biophysical Journal

View full text Add to dashboard Cite

Considerable mechanistic insight has been gained into amyloid aggregation; however, a large class of non-amyloid protein aggregates are considered "amorphous," and in most cases little is known about their mechanisms. Amorphous aggregation of γ-crystallins in the eye lens causes a widespread disease of aging, cataract. We combined simulations and experiments to study the mechanism of aggregation of two γD-crystallin mutants, W42R and W42Q -the former a congenital cataract mutation, and the latter a mimic of age-related oxidative damage. We found that formation of an internal disulfide was necessary and sufficient for aggregation under physiological conditions. Twochain all-atom simulations predicted that one nonnative disulfide in particular, between Cys32 and Cys41, was likely to stabilize an unfolding intermediate prone to intermolecular interactions. Mass spectrometry and mutagenesis experiments confirmed the presence of this bond in the aggregates and its necessity for oxidative aggregation under physiological conditions in vitro. Mining the simulation data linked formation of this disulfide to extrusion of the N-terminal β-hairpin and rearrangement of the native β-sheet topology.

show abstract

The Mitochondrial Intermembrane Space Oxireductase Mia40 Funnels the Oxidative Folding Pathway of the Cytochrome c Oxidase Assembly Protein Cox19

Cited by 16 publications

References 31 publications

Conformational Characterization and Classification of Intrinsically Disordered Proteins by Native Mass Spectrometry and Charge‐State Distribution Analysis

Conformational Characterization and Classification of Intrinsically Disordered Proteins by Native Mass Spectrometry and Charge‐State Distribution Analysis

Disulfide driven folding for a conditionally disordered protein

An Internal Disulfide Locks a Misfolded Aggregation-Prone Intermediate in Cataract-Linked Mutants of Human Gamma-D Crystallin

Contact Info

Product

Resources

About