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Formation and transfer of disulphide bonds in living cells

Nature Reviews Molecular Cell Biology volume 3, pages 836–847 (2002)Cite this article

Key Points

  • The formation of structural disulphide bonds in cellular proteins is a catalysed process that involves many proteins and small molecules. The primary pathways of disulphide-bond formation are localized in the endoplasmic reticulum (ER) of eukaryotic cells and the periplasmic space of prokaryotic cells.

  • The core pathways that promote disulphide-bond formation in prokaryotes and eukaryotes share many similarities. Both pathways include soluble thiol-disulphide oxidoreductases that donate disulphide bonds directly to substrate proteins, as well as membrane-associated enzymes that maintain the soluble enzymes in a redox-active form.

  • Protein oxidation in the ER relies on the membrane-associated proteins Ero1 (ER oxidoreductin) and Erv2, and the soluble thiol-disulphide oxidoreductase protein disulphide isomerase (PDI). The prokaryotic protein oxidation system uses the integral membrane protein DsbB and the soluble enzyme DsbA.

  • In addition to the DsbA–DsbB pathway for disulphide-bond formation, prokaryotes also contain a pathway for the isomerization of non-native disulphide bonds. This pathway includes the membrane protein DsbD and the soluble enzyme DsbC. At present, a reduction pathway similar to the DsbC–DsbD pathway has not been characterized in eukaryotes.

  • The protein oxidation and isomerization pathways in prokaryotes and eukaryotes use a conserved thiol-disulphide exchange mechanism to transfer disulphide bonds between components. In addition to these inter-protein transfer events, several of the enzymes also seem to catalyse the intra-protein transfer of disulphide bonds between their own cysteine pairs.

  • The bacterial DsbA–DsbB protein oxidation system is driven by oxidizing equivalents derived from the cellular respiratory electron-transport chain. The source of oxidizing equivalents for ER protein oxidation is not as well characterized. Flavin moieties seem to provide a source of oxidizing equivalents, but the sources for flavin oxidation are not well understood.

  • The identification of enzymatic pathways of disulphide-bond formation has raised many questions about the role of the principal cellular small-molecule redox compound glutathione. Glutathione was originally believed to drive protein oxidation; however, more recent experiments show that glutathione is not required for oxidative protein folding. Instead, it has been suggested that glutathione functions as a net reductant in the ER, perhaps protecting the ER under hyperoxidizing conditions.

Abstract

Protein disulphide bonds are formed in the endoplasmic reticulum of eukaryotic cells and the periplasmic space of prokaryotic cells. The main pathways that catalyse the formation of protein disulphide bonds in prokaryotes and eukaryotes are remarkably similar, and they share several mechanistic features. The recent identification of new redox-active proteins in humans and yeast that mechanistically parallel the more established redox-active enzymes indicates that there might be further uncharacterized redox pathways throughout the cell.

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Figure 1: Pathways for protein oxidation in the endoplasmic reticulum of Saccharomyces cerevisiae.
Figure 2: Periplasmic pathways for protein oxidation and isomerization in Escherichia coli.

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Authors and Affiliations

  1. Department of Biology, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts, USA

    Carolyn S. Sevier & Chris A. Kaiser

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  1. Carolyn S. Sevier
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  2. Chris A. Kaiser
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Correspondence to Chris A. Kaiser.

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DATABASES

Pfam

thioredoxin

<i>Saccharomyces</i> Genome Database

GSH1

Swiss-Prot

DsbA

DsbB

DsbC

DsbD

Ero1

Erv1

Erv2

Eug1

Mpd1

Mpd2

PDI

Glossary

THIOL-REDOX REACTION

A reaction that involves the transfer of electrons from a donor molecule to an acceptor molecule if one of the molecules is a thiol-containing compound.

THIOL-DISULPHIDE EXCHANGE REACTION

A thiol-redox reaction that involves the exchange of electrons between a compound with free thiols and a disulphide-bonded molecule, which results in the transfer of a disulphide bond from one molecule to another.

GLUTATHIONE

A tripeptide — composed of glutamic acid, cysteine and glycine — that is the principal small thiol-containing molecule in the cell.

THIOL-DISULPHIDE OXIDOREDUCTASE

An enzyme that catalyses the transfer of electrons or hydrogen between molecules.

THIOREDOXIN

A ubiquitous small soluble protein with redox-active cysteines that catalyses thiol-disulphide exchange reactions.

PROTEIN DISULPHIDE ISOMERASE

A soluble protein with two thioredoxin-like domains that each contain a redox-active cysteine pair that donates disulphide bonds to newly synthesized proteins in the eukaryotic ER.

REDOX (REDUCTION-OXIDATION) POTENTIAL

The propensity of a given protein (or molecule) to gain or donate electrons, which is usually expressed as an electrochemical potential in volts. A protein's redox potential can be measured by quantifying the steady-state ratios of the reduced and oxidized forms of this protein that are present in a buffer of defined redox composition. The term 'reduction potential' is often used instead.

OXIDIZING EQUIVALENTS

The loss of electrons by a molecule (this equals the gain of oxidizing equivalents).

MIXED-DISULPHIDE BOND

A disulphide bond that is formed between two proteins or redox molecules. These bonds are often transient and reflect an intermediate in the transfer of oxidizing equivalents between redox-active proteins and molecules.

QUINONES

A group of lipid-soluble compounds that function as electron carriers in the electron-transport chain reactions of cellular respiration.

ERV-LIKE PROTEIN FAMILY

A family of flavoprotein thiol-oxidases — named after their homology to the yeast protein Erv1 — that couples the oxidation of free thiols with the reduction of molecular oxygen to hydrogen peroxide.

(K/H)DEL SIGNAL

An ER-localization motif for soluble lumenal proteins that includes the short carboxy-terminal sequence Lys/His-Asp-Glu-Leu.

CHAPERONE

A protein that catalyses the correct folding of newly synthesized or denatured proteins into their native conformations.

RESPIRATORY ELECTRON-TRANSPORT CHAIN

A series of redox-active membrane proteins and small molecules in either the bacterial plasma membrane or the mitochondrial inner membrane that carry out the step-by-step transfer of electrons from NADH and FADH2 to O2 with the concomitant generation of a membrane proton potential.

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Sevier, C., Kaiser, C. Formation and transfer of disulphide bonds in living cells. Nat Rev Mol Cell Biol 3, 836–847 (2002). https://doi.org/10.1038/nrm954

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