Formation and transfer of disulphide bonds in living cells

Sevier, Carolyn S.; Kaiser, Chris A.

doi:10.1038/nrm954

Cited by 681 publications

(570 citation statements)

References 125 publications

Supporting

Mentioning

551

Contrasting

Unclassified

Order By: Relevance

“…This is currently an issue because the natural redox partner(s) of yeast Erv2p have yet to be identified. Although crosslinking experiments suggest that PDI1p interacts with Erv2p (7,49), in vitro experiments with reduced PDI1p show that it is a poor substrate of this oxidase (24). This kinetic sluggishness has led to the suggestion that other ER redox proteins may mediate transfer of reducing equivalents from client unfolded proteins to Erv2p (24).…”

Section: The Erv/alr Family Of Sulfhydryl Oxidases: a Comparisonmentioning

confidence: 99%

Erv2p: Characterization of the Redox Behavior of a Yeast Sulfhydryl Oxidase

2007

View full text Add to dashboard Cite

The FAD prosthetic group of the ERV/ALR family of sulfhydryl oxidases is housed at the mouth of a 4-helix bundle and communicates with a pair of juxtaposed cysteine residues that form the proximal redox active disulfide. Most of these enzymes have one or more additional distal disulfide redox centers that facilitate the transfer of reducing equivalents from the dithiol substrates of these oxidases to the isoalloxazine ring where the reaction with molecular oxygen occurs. The present study examines yeast Erv2p and compares the redox behavior of this ER luminal protein with the augmenter of liver regeneration, a sulfhydryl oxidase of the mitochondrial intermembrane space, and a larger protein containing the ERV/ALR domain, quiescin-sulfhydryl oxidase (QSOX). Dithionite and photochemical reductions of Erv2p show full reduction of the flavin cofactor after the addition of 4-electrons with a mid-point potential of -200 mV at pH 7.5. A charge-transfer complex between a proximal thiolate and the oxidized flavin is not observed in Erv2p consistent with a distribution of reducing equivalents over the flavin and distal disulfide redox centers. Upon coordination with Zn 2+ , full reduction of Erv2p requires 6-electrons. Zn 2+ also strongly inhibits Erv2p when assayed using tris(2-carboxyethyl)phosphine (TCEP) as the reducing substrate of the oxidase. In contrast to QSOX, Erv2p shows a comparatively low turnover with a range of small thiol substrates, with reduced Escherichia coli thioredoxin and with unfolded proteins. Rapid reaction studies confirm that reduction of the flavin center of Erv2p is rate-limiting during turnover with molecular oxygen. This comparison of the redox properties between members of the ERV/ALR family of sulfhydryl oxidases provides insights into their likely roles in oxidative protein folding.Over the last decade a number of eukaryotic flavin-dependent sulfhydryl oxidases have been described that catalyze the net generation of disulfide bonds:They include: yeast ERV1 (essential for respiration and vegetative growth) (1) and its mammalian counterpart augmenter of liver regeneration, ALR, (2-4); yeast Erv2p (5-7); an Arabidopsis homolog of these proteins (8,9); yeast and its vertebrate orthologs Ero1α and Ero1β (15,16); and finally larger multidomain sulfhydryl oxidases, containing an ERV/ALR module fused to a redox active thioredoxin domain, that are found principally in multicellular organisms (17)(18)(19)(20) The ERV/ALR proteins and their larger cousins in the QSOX family all contain a diminutive helix-rich flavin binding domain first reported for yeast Erv2p by Fass and coworkers (6) and for rat ALR by Rose and colleagues (4). Subunit A of the Erv2p homodimer is shown in the foreground of Figure 1. The isoalloxazine ring is inserted at the end of a bundle of four helices in the A subunit with its C2/N3 region exposed to solvent (6). The proximal disulfide C121-124 is placed so that the sulfur of C124 is adjacent (3.4 Å) to the C 4a position of the flavin. This locus is consistent with i...

show abstract

Section: The Erv/alr Family Of Sulfhydryl Oxidases: a Comparisonmentioning

confidence: 99%

Erv2p: Characterization of the Redox Behavior of a Yeast Sulfhydryl Oxidase

2007

View full text Add to dashboard Cite

show abstract

“…mined 7 . The redox gradient between the ER and the cytosol seems to be important for intercompartmental signalling, particularly in the integrated response to oxidative stress, in which adaptive responses emanating from different compartments are coordinated 9 .…”

Section: Er-associated Degradationmentioning

confidence: 99%

“…The requirement for oxidative equivalents is fulfilled by Ero1 proteins, which transfer electrons from protein disulphide isomerase (PDI) to oxygen through a series of specific interchange reactions 7,8 The endoplasmic reticulum (ER) is a factory where secretory proteins are manufactured, and where stringent quality-control systems ensure that only correctly folded proteins are sent to their final destinations. The changing needs of the ER factory are monitored by integrated signalling pathways that constantly adjust the levels of folding assistants.…”

Section: Disulphide-bond Formationmentioning

confidence: 99%

Quality control in the endoplasmic reticulum protein factory

Sitia

Braakman

2003

Nature

608

433

View full text Add to dashboard Cite

“…Erv1p belongs to the class of FAD-containing sulfhydryl oxidases which transfer electrons from thiols to non-thiol electron acceptors and thereby generate disulfide bonds in proteins (28,29). Indeed, thiol oxidase activity has been measured for Erv1p in vitro (30 -32).…”

mentioning

confidence: 99%

Functional Characterization of Mia40p, the Central Component of the Disulfide Relay System of the Mitochondrial Intermembrane Space

Grumbt

Stroobant

Terziyska

et al. 2007

Journal of Biological Chemistry

116

View full text Add to dashboard Cite

Mia40p and Erv1p are components of a translocation pathway for the import of cysteine-rich proteins into the intermembrane space of mitochondria. We have characterized the redox behavior of Mia40p and reconstituted the disulfide transfer system of Mia40p by using recombinant functional C-terminal fragment of Mia40p, Mia40C, and Erv1p. Oxidized Mia40p contains three intramolecular disulfide bonds. One disulfide bond connects the first two cysteine residues in the CPC motif. The second and the third bonds belong to the twin CX 9 C motif and bridge the cysteine residues of two CX 9 C segments. In contrast to the stabilizing disulfide bonds of the twin CX 9 C motif, the first disulfide bond was easily accessible to reducing agents. Partially reduced Mia40C generated by opening of this bond as well as fully reduced Mia40C were oxidized by Erv1p in vitro. In the course of this reaction, mixed disulfides of Mia40C and Erv1p were formed. Reoxidation of fully reduced Mia40C required the presence of the first two cysteine residues in Mia40C. However, efficient reoxidation of a Mia40C variant containing only the cysteine residues of the twin CX 9 C motif was observed when in addition to Erv1p low amounts of wild type Mia40C were present. In the reconstituted system the thiol oxidase Erv1p was sufficient to transfer disulfide bonds to Mia40C, which then could oxidize the variant of Mia40C. In summary, we reconstituted a disulfide relay system consisting of Mia40C and Erv1p.

show abstract

Formation and transfer of disulphide bonds in living cells

Cited by 681 publications

References 125 publications

Erv2p: Characterization of the Redox Behavior of a Yeast Sulfhydryl Oxidase

Erv2p: Characterization of the Redox Behavior of a Yeast Sulfhydryl Oxidase

Quality control in the endoplasmic reticulum protein factory

Functional Characterization of Mia40p, the Central Component of the Disulfide Relay System of the Mitochondrial Intermembrane Space

Contact Info

Product

Resources

About