Protein Disulfide Isomerases

Hiram F Gilbert, Baylor College of Medicine, Houston, Texas, USA

Published online: April 2011

DOI: 10.1002/9780470015902.a0003021.pub2

Protein disulfide isomerase (PDI) is a multifunctional protein that facilitates the formation of correct disulfide crosslinks between cysteine residues during the early stages of protein folding and secretion in the endoplasmic reticulum. It is a member of a large family of oxidoreductases that catalyse exchange reactions between thiols and disulfides. PDI is a multidomain protein, consisting of four tandem thioredoxin domains. The N- and C-terminal thioredoxin domains have catalytic disulfide/dithiol centers while the two internal domains are structural and provide additional interactions with the protein substrates. The catalysis of disulfide formation relies almost entirely on the high reactivity of PDI's active site disulfides. However, the ability to catalyse disulfide isomerisation requires multiple domains. In the cell, PDI's essential activity is the formation of disulfide bonds, but it does catalyse isomerisation.

Key Concepts:

  • PDI structure consists of four tandem thiredoxin domains.
  • The ability to form disulfides between cysteines in substrate proteins results from highly reactive active site disulfides.
  • PDI can correct (isomerise) misformed disulfides by reducing incorrect disulfides and reoxidising in a different configuration.
  • The multidomain structure is needed for catalysis of isomerisation but a single catalytic domain is sufficient to catalyse substrate oxidation.
  • In the cell, the essential activity of PDI is its ability to form substrate disulfides.

Keywords: PDI; disulfide; protein folding; chaperone; disulfide isomerisation

Figure 1. Three-dimensional structures of the active site of the a domain of PDI. Arrows point to the more N-terminal cysteine residue of each active site (nucleophilic cysteine, CGHC). The structure was drawn using the Swiss-Pdb Viewer (Guex and Peitsch, 1997) using the coordinates in PDB file 2B5E (Tian et al., 2006).
Figure 2. The domain structure of PDI. The active site cysteines in the a and a¢ domains are shown as space-filling models. The structure was drawn using the Swiss-Pdb Viewer (Guex and Peitsch, 1997) using the coordinates in PDB file 2B5E (Tian et al., 2006).
Figure 3. Catalysis of disulfide isomerisation. Disulfide isomerisation is initiated by attack of the nucleophilic cysteine of the PDI active site (CGHC) on a substrate disulfide. The substrate-PDI covalent complex can be resolved by two possible isomerisation mechanisms. Which pathway occurs is determined by the action of the resolving cysteine (CGHC) at the PDI active site. If an intramolecular rearrangement of the substrate disulfides occurs slowly, the resolving cysteine will displace the substrate mixed disulfide from the nucleophilic cysteine (CGHC) reducing it and forming an oxidised PDI active site. Further cycles of reduction/reoxidation in different disulfide configurations would result in overall isomerisation. Adapted from Wilkinson and Gilbert (2004), with permission from Elsevier.
Figure 4. Redox balance in the endoplasmic reticulum. Oxidising equivalents are provided to the eukaryotic ER by the oxidase Ero1, which oxidises the PDI active site. Reducing equivalents, to help balance the redox state and allow disulfide isomerisation are introduced from the cytoplasm by translocation of the newly synthesises substrate and by a glutathione (GSH)-dependent mechanism.
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 References
    Cuozzo JW and Kaiser CA (1999) Competition between glutathione and protein thiols for disulphide-bond formation. Nature Cell Biology 1: 130–135.
    Darby NJ, Penka E and Vincentelli R (1998) The multi-domain structure of protein disulfide isomerase is essential for high catalytic efficiency. Journal of Molecular Biology 276: 239–247.
    Ellgaard L and Ruddock LW (2005) The human protein disulphide isomerase family: substrate interactions and functional properties. EMBO Report 6: 28–32.
    Frand AR and Kaiser CA (1998) The ERO1 gene of yeast is required for oxidation of protein dithiols in the endoplasmic reticulum. Molecular Cell 1: 161–170.
    Gilbert HF (1998) Protein disulfide isomerase. Methods in Enzymology 290: 26–50.
    Guex N and Peitsch MC (1997) SWISS-MODEL and the Swiss-PdbViewer: an environment for comparative protein modeling. Electrophoresis 18: 2714–2723.
    Hatahet F and Ruddock LW (2009) Protein disulfide isomerase: a critical evaluation of its function in disulfide bond formation. Antioxidants & Redox Signaling 11: 2807–2850.
    Holmgren A (1989) Thioredoxin and glutaredoxin systems. Journal of Biological Chemitry 25; 264: 13963–13966.
    Kadokura H and Beckwith J (2010) Mechanisms of oxidative protein folding in the bacterial cell envelope. Antioxid Redox Signal 13: 1231–1246.
    Kivirikko KI and Pihlajaniemi T (1998) Collagen hydroxylases and the protein disulfide isomerase subunit of prolyl 4-hydroxylases. Advances in Enzymology and Related Areas of Molecular Biology 72: 325–398.
    Klappa P, Ruddock LW, Darby NJ and Freedman RB (1998) The b¢ domain provides the principal peptide-binding site of protein disulfide isomerase but all domains contribute to binding of misfolded proteins. EMBO Journal 17: 927–935.
    Kulp MS, Frickel E-M, Ellgaard L and Weissman JS (2006) Domain architecture of protein-disulfide isomerase facilitates its dual role as an oxidase and an isomerase in Ero1p-mediated disulfide formation. Journal of Biological Chemistry 281: 876–884.
    Lillig CH and Holmgren A (2007) Thioredoxin and related molecules: from biology to health and disease. Antioxid Redox Signal 9: 25–47.
    Lu J and Holmgren A (2009) Selenoproteins. Journal of Biological Chemistry 284: 723–727.
    Lyles MM and Gilbert HF (1991) Catalysis of the oxidative folding of ribonuclease A by protein disulfide isomerase: dependence of the rate on the composition of the redox buffer. Biochemistry 30: 613–619.
    Lyles MM and Gilbert HF (1994) Mutations in the thioredoxin sites of protein disulfide isomerase reveal functional non-equivalence of the N- and C-terminal domains. Journal of Biological Chemistry 269: 30946–30952.
    Martin JL, Bardwell JCA and Kuriyan J (1993) Crystal structure of the DsbA protein required for disulphide bond formation in vivo. Nature 365: 464–468.
    Merksamer PI, Trusina A and Papa FR (2008) Real-time redox measurements during endoplasmic reticulum stress reveal interlinked protein folding functions. Cell 135: 933–847.
    Pollard MG, Travers KJ and Weissman JS (1998) ERO1p: a novel and ubiquitous protein with an essential role in oxidative protein folding in the endoplasmic reticulum. Molecular Cell 1: 171–182.
    Primm TP, Walker KW and Gilbert HF (1996) Facilitated protein aggregation. Effects of calcium on the chaperone and anti-chaperone activity of protein disulfide-isomerase. Journal of Biological Chemistry 271: 33664–33669.
    Raina S and Missiakas D (1997) Making and breaking disulfide bonds. Annual Review of Microbiology 51: 179–202.
    Rietsch A and Beckwith J (1998) The genetics of disulfide bond metabolism. Annual Review of Genetics 32: 163–184.
    Schwaller MF, Wilkinson B and Gilbert HF (2003) Reduction/reoxidation cycles contribute to catalysis of disulfide isomerisation by protein disulfide isomerase. Journal of Biological Chemistry 278: 7154–7159.
    Sideraki V and Gilbert HF (2000) Mechanism of the antichaperone activity of protein disulfide isomerase: facilitated assembly of large, insoluble aggregates of denatured lysozyme and PDI. Biochemistry 39: 1180–1188.
    Solovyov A, Xiao R and Gilbert HF (2004) Sulfhydryl oxidation, not disulfide isomerisation, is the principal function of protein disulfide isomerase in yeast Saccharomyces cerevisiae. Journal of Biological Chemistry 279: 34095–34100.
    Tian G, Kober F-X, Lewandrowski U et al. (2008) The catalytic activity of protein disulfide isomerase requires a conformationally flexible molecule. Journal of Biological Chemistry 283: 33630–33640.
    Tian G, Xiang S, Noiva R, Lennarz WJ and Schindelin H (2006) The crystal structure of yeast protein disulfide isomerase suggests cooperativity between its active sites. Cell 124: 61–73.
    Walker KW and Gilbert HF (1995) Oxidation of kinetically trapped thiols by protein disulfide isomerase. Biochemistry 34: 13642–13650.
    book Walker KW and Gilbert HF (1997a) "Protein disulfide isomerase". In: Fink A and Goto H (eds) Structure and Function of Molecular Chaperones: The Role of Chaperones in the Life Cycle of Proteins, pp. 331–360. New York: Marcel Dekker.
    Walker KW and Gilbert HF (1997b) Scanning and escape during protein-disulfide isomerase-assisted protein folding. Journal of Biological Chemistry 272: 8845–8848.
    Wang CC and Tsou CL (1998) Enzymes as chaperones and chaperones as enzymes. FEBS Letters 425: 382–384.
    Wang L, Fast DG and Attie AD (1997) The enzymatic and non-enzymatic roles of protein-disulfide isomerase in apoliprotein B secretion. Journal of Biological Chemistry 272: 27644–27651.
    Westphal V, Spetzler JC, Meldal M, Christensen LL and Winther JR (1998) Kinetic analysis of the mechanism and specificity of protein-disulfide isomerase using fluorescence-quenched peptides. Journal of Biological Chemistry 273: 24992–24999.
    Whitley EM, Hsu TA and Betenbaugh MJ (1997) Thioredoxin domain non-equivalence and anti-chaperone activity of protein disulfide isomerase mutants in vivo. Journal of Biological Chemistry 272: 22556–22563.
    Wilkinson B and Gilbert HF (2004) Protein disulfide isomerase. Biochemica et Biophysica Acta 1699: 35–44.
    Xiao R, Wilkinson B, Solovyov A et al. (2004) Protein disulfide isomerase is an oxidase and isomerase in the S. cerevisiae endoplasmic reticulum. Journal of Biological Chemistry 279: 49780–49786.
 Further Reading
    Braakman I (2009) Entering a new era with ero. Nature Reviews. Molecular Cell Biology 10: 503.
    Freedman RB, Hawkins HC and McLaughlin SH (1995) Protein disulfide isomerase. Methods in Enzymology 251: 397–406.
    Gorres KL and Raines RT (2010) Prolyl 4-hydroxylase. Critical Review of Biochemistry and Molecular Biology 45: 106–124.
    Hatahet F and Ruddock LW (2007) Substrate recognition by the protein disulfide isomerases. FEBS Journal 4: 5223–5234.
    Jensen KS, Hansen RE and Winther JR (2009) Kinetic and thermodynamic aspects of cellular thiol-disulfide redox regulation. Antioxidants and Redox Signaling 11: 1047–1058.
    Mamathambika BS and Bardwell JC (2008) Disulfide-linked protein folding pathways. Annual Review of Cell and Developmental Biology 24: 211–235.

Related Sub-Topics:

Protein Structure | Protein Folding, Evolution and Degradation | Biomolecular Interactions | Proteins: Structure, Function, Metabolism | Enzymes: Structure and Action Mechanism
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Article Title: Protein Disulfide Isomerases
 
How to Cite close
Gilbert, Hiram F(Apr 2011) Protein Disulfide Isomerases. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0003021.pub2]