Substrate-Induced Unfolding of Protein Disulfide Isomerase Displaces the Cholera Toxin A1 Subunit from Its Holotoxin

English version České info

To generate a cytopathic effect, the catalytic A1 subunit of cholera toxin (CT) must be separated from the rest of the toxin. Protein disulfide isomerase (PDI) is thought to mediate CT disassembly by acting as a redox-driven chaperone that actively unfolds the CTA1 subunit. Here, we show that PDI itself unfolds upon contact with CTA1. The substrate-induced unfolding of PDI provides a novel molecular mechanism for holotoxin disassembly: we postulate the expanded hydrodynamic radius of unfolded PDI acts as a wedge to dislodge reduced CTA1 from its holotoxin. The oxidoreductase activity of PDI was not required for CT disassembly, but CTA1 displacement did not occur when PDI was locked in a folded conformation or when its substrate-induced unfolding was blocked due to the loss of chaperone function. Two other oxidoreductases (ERp57 and ERp72) did not unfold in the presence of CTA1 and did not displace reduced CTA1 from its holotoxin. Our data establish a new functional property of PDI that may be linked to its role as a chaperone that prevents protein aggregation.

Vyšlo v časopise: Substrate-Induced Unfolding of Protein Disulfide Isomerase Displaces the Cholera Toxin A1 Subunit from Its Holotoxin. PLoS Pathog 10(2): e32767. doi:10.1371/journal.ppat.1003925
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1003925

Souhrn

Zdroje

1. HatahetF, RuddockLW (2007) Substrate recognition by the protein disulfide isomerases. FEBS J 274: 5223–5234.

2. KozlovG, MaattanenP, ThomasDY, GehringK (2010) A structural overview of the PDI family of proteins. FEBS J 277: 3924–3936.

3. WilkinsonB, GilbertHF (2004) Protein disulfide isomerase. Biochim Biophys Acta 1699: 35–44.

4. TianG, KoberFX, LewandrowskiU, SickmannA, LennarzWJ, et al. (2008) The catalytic activity of protein-disulfide isomerase requires a conformationally flexible molecule. J Biol Chem 283: 33630–33640.

5. NakasakoM, MaenoA, KurimotoE, HaradaT, YamaguchiY, et al. (2010) Redox-dependent domain rearrangement of protein disulfide isomerase from a thermophilic fungus. Biochemistry 49: 6953–6962.

6. ServeO, KamiyaY, MaenoA, NakanoM, MurakamiC, et al. (2010) Redox-dependent domain rearrangement of protein disulfide isomerase coupled with exposure of its substrate-binding hydrophobic surface. J Mol Biol 396: 361–374.

7. TaylorM, BanerjeeT, RayS, TatulianSA, TeterK (2011) Protein disulfide isomerase displaces the cholera toxin A1 subunit from the holotoxin without unfolding the A1 subunit. J Biol Chem 286: 22090–22100.

8. TsaiB, RodighieroC, LencerWI, RapoportTA (2001) Protein disulfide isomerase acts as a redox-dependent chaperone to unfold cholera toxin. Cell 104: 937–948.

9. FuX, ZhuBT (2009) Human pancreas-specific protein disulfide isomerase homolog (PDIp) is redox-regulated through formation of an inter-subunit disulfide bond. Arch Biochem Biophys 485: 1–9.

10. WangC, YuJ, HuoL, WangL, FengW, et al. (2012) Human protein-disulfide isomerase is a redox-regulated chaperone activated by oxidation of domain a'. J Biol Chem 287: 1139–1149.

11. FuXM, ZhuBT (2010) Human pancreas-specific protein disulfide-isomerase (PDIp) can function as a chaperone independently of its enzymatic activity by forming stable complexes with denatured substrate proteins. Biochem J 429: 157–169.

12. QuanH, FanG, WangCC (1995) Independence of the chaperone activity of protein disulfide isomerase from its thioredoxin-like active site. J Biol Chem 270: 17078–17080.

13. SongJL, WangCC (1995) Chaperone-like activity of protein disulfide-isomerase in the refolding of rhodanese. Eur J Biochem 231: 312–316.

14. PrimmTP, WalkerKW, GilbertHF (1996) Facilitated protein aggregation. Effects of calcium on the chaperone and anti-chaperone activity of protein disulfide-isomerase. J Biol Chem 271: 33664–33669.

15. UeharaT, NakamuraT, YaoD, ShiZQ, GuZ, et al. (2006) S-nitrosylated protein-disulphide isomerase links protein misfolding to neurodegeneration. Nature 441: 513–517.

16. ChengH, WangL, WangCC (2010) Domain a' of protein disulfide isomerase plays key role in inhibiting alpha-synuclein fibril formation. Cell Stress Chaperones 15: 415–421.

17. SmithWW, JiangH, PeiZ, TanakaY, MoritaH, et al. (2005) Endoplasmic reticulum stress and mitochondrial cell death pathways mediate A53T mutant alpha-synuclein-induced toxicity. Hum Mol Genet 14: 3801–3811.

18. CaiH, WangCC, TsouCL (1994) Chaperone-like activity of protein disulfide isomerase in the refolding of a protein with no disulfide bonds. J Biol Chem 269: 24550–24552.

19. SanchezJ, HolmgrenJ (2008) Cholera toxin structure, gene regulation and pathophysiological and immunological aspects. Cell Mol Life Sci 65: 1347–1360.

20. WernickNLB, ChinnapenDJ-F, ChoJA, LencerWI (2010) Cholera toxin: an intracellular journey into the cytosol by way of the endoplasmic reticulum. Toxins 2: 310–325.

21. BanerjeeT, PandeA, JoblingMG, TaylorM, MasseyS, et al. (2010) Contribution of subdomain structure to the thermal stability of the cholera toxin A1 subunit. Biochemistry 49: 8839–8846.

22. MasseyS, BanerjeeT, PandeAH, TaylorM, TatulianSA, et al. (2009) Stabilization of the tertiary structure of the cholera toxin A1 subunit inhibits toxin dislocation and cellular intoxication. J Mol Biol 393: 1083–1096.

23. TaylorM, BanerjeeT, Navarro-GarciaF, HuertaJ, MasseyS, et al. (2011) A therapeutic chemical chaperone inhibits cholera intoxication and unfolding/translocation of the cholera toxin A1 subunit. PLoS ONE 6: e18825.

24. PandeAH, ScaglioneP, TaylorM, NemecKN, TuthillS, et al. (2007) Conformational instability of the cholera toxin A1 polypeptide. J Mol Biol 374: 1114–1128.

25. AmpapathiRS, CreathAL, LouDI, CraftJWJr, BlankeSR, et al. (2008) Order-disorder-order transitions mediate the activation of cholera toxin. J Mol Biol 377: 748–760.

26. RodighieroC, TsaiB, RapoportTA, LencerWI (2002) Role of ubiquitination in retro-translocation of cholera toxin and escape of cytosolic degradation. EMBO Rep 3: 1222–1227.

27. RayS, TaylorM, BanerjeeT, TatulianSA, TeterK (2012) Lipid rafts alter the stability and activity of the cholera toxin A1 subunit. J Biol Chem 287: 30395–30405.

28. MajoulI, FerrariD, SolingHD (1997) Reduction of protein disulfide bonds in an oxidizing environment. The disulfide bridge of cholera toxin A-subunit is reduced in the endoplasmic reticulum. FEBS Lett 401: 104–108.

29. MekalanosJJ, CollierRJ, RomigWR (1979) Enzymic activity of cholera toxin. II. Relationships to proteolytic processing, disulfide bond reduction, and subunit composition. J Biol Chem 254: 5855–5861.

30. O'NealCJ, JoblingMG, HolmesRK, HolWG (2005) Structural basis for the activation of cholera toxin by human ARF6-GTP. Science 309: 1093–1096.

31. ZhangRG, ScottDL, WestbrookML, NanceS, SpanglerBD, et al. (1995) The three-dimensional crystal structure of cholera toxin. J Mol Biol 251: 563–573.

32. TatulianSA (2010) Structural analysis of proteins by isotope-edited FTIR spectroscopy. Spectroscopy Int J 24: 37–43.

33. DecaturSM (2006) Elucidation of residue-level structure and dynamics of polypeptides via isotope-edited infrared spectroscopy. Acc Chem Res 39: 169–175.

34. HarisPI (2013) Probing protein-protein interaction in biomembranes using Fourier transform infrared spectroscopy. Biochim Biophys Acta 1828: 2265–2271.

35. TianG, XiangS, NoivaR, LennarzWJ, SchindelinH (2006) The crystal structure of yeast protein disulfide isomerase suggests cooperativity between its active sites. Cell 124: 61–73.

36. GoinsB, FreireE (1988) Thermal stability and intersubunit interactions of cholera toxin in solution and in association with its cell-surface receptor ganglioside GM1. Biochemistry 27: 2046–2052.

37. EnamiI, KamoM, OhtaH, TakahashiS, MiuraT, et al. (1998) Intramolecular cross-linking of the extrinsic 33-kDa protein leads to loss of oxygen evolution but not its ability of binding to photosystem II and stabilization of the manganese cluster. J Biol Chem 273: 4629–4634.

38. NovakP, KruppaGH (2008) Intra-molecular cross-linking of acidic residues for protein structure studies. Eur J Mass Spectrom (Chichester, Eng) 14: 355–365.

39. WernickNL, De LucaH, KamWR, LencerWI (2010) N-terminal extension of the cholera toxin A1-chain causes rapid degradation after retrotranslocation from endoplasmic reticulum to cytosol. J Biol Chem 285: 6145–6152.

40. DickerhofN, KleffmannT, JackR, McCormickS (2011) Bacitracin inhibits the reductive activity of protein disulfide isomerase by disulfide bond formation with free cysteines in the substrate-binding domain. FEBS J 278: 2034–2043.

41. HoribeT, NagaiH, SakakibaraK, HagiwaraY, KikuchiM (2001) Ribostamycin inhibits the chaperone activity of protein disulfide isomerase. Biochem Biophys Res Commun 289: 967–972.

42. WalkerAK, FargMA, ByeCR, McLeanCA, HorneMK, et al. (2010) Protein disulphide isomerase protects against protein aggregation and is S-nitrosylated in amyotrophic lateral sclerosis. Brain 133: 105–116.

43. TeterK, JoblingMG, SentzD, HolmesRK (2006) The cholera toxin A13 subdomain is essential for interaction with ADP-ribosylation factor 6 and full toxic activity but is not required for translocation from the endoplasmic reticulum to the cytosol. Infect Immun 74: 2259–2267.

44. OrlandiPA (1997) Protein-disulfide isomerase-mediated reduction of the A subunit of cholera toxin in a human intestinal cell line. J Biol Chem 272: 4591–4599.

45. LencerWI, de AlmeidaJB, MoeS, StowJL, AusielloDA, et al. (1993) Entry of cholera toxin into polarized human intestinal epithelial cells. Identification of an early brefeldin A sensitive event required for A1-peptide generation. J Clin Invest 92: 2941–2951.

46. SchelhaasM, MalmstromJ, PelkmansL, HaugstetterJ, EllgaardL, et al. (2007) Simian Virus 40 depends on ER protein folding and quality control factors for entry into host cells. Cell 131: 516–529.

47. LencerWI, TsaiB (2003) The intracellular voyage of cholera toxin: going retro. Trends Biochem Sci 28: 639–645.

48. WangC, ChenS, WangX, WangL, WallisAK, et al. (2010) Plasticity of human protein disulfide isomerase: evidence for mobility around the X-linker region and its functional significance. J Biol Chem 285: 26788–26797.

49. BellisolaG, FracassoG, IppolitiR, MenestrinaG, RosenA, et al. (2004) Reductive activation of ricin and ricin A-chain immunotoxins by protein disulfide isomerase and thioredoxin reductase. Biochem Pharmacol 67: 1721–1731.

50. SpoonerRA, WatsonPD, MarsdenCJ, SmithDC, MooreKA, et al. (2004) Protein disulphide-isomerase reduces ricin to its A and B chains in the endoplasmic reticulum. Biochem J 383: 285–293.

51. McKeeML, FitzGeraldDJ (1999) Reduction of furin-nicked Pseudomonas exotoxin A: an unfolding story. Biochemistry 38: 16507–16513.

52. HolmesRK, TwiddyEM (1983) Characterization of monoclonal antibodies that react with unique and cross-reacting determinants of cholera enterotoxin and its subunits. Infect Immun 42: 914–923.

53. NguyenVD, SaaranenMJ, KaralaAR, LappiAK, WangL, et al. (2011) Two endoplasmic reticulum PDI peroxidases increase the efficiency of the use of peroxide during disulfide bond formation. J Mol Biol 406: 503–515.

54. Tatulian SA (2013) Structural characterization of membrane proteins and peptides by FTIR and ATR-FTIR spectroscopy. In: Kleinschmidt JH, editor. Lipid-protein interactions: methods and protocols. New York: Humana Press. pp. 177–218.

55. TeterK, JoblingMG, HolmesRK (2004) Vesicular transport is not required for the cytoplasmic pool of cholera toxin to interact with the stimulatory alpha subunit of the heterotrimeric G protein. Infect Immun 72: 6826–6835.

56. TaylorM, BanerjeeT, VanBennekomN, TeterK (2012) Detection of toxin translocation into the host cytosol by surface plasmon resonance. Journal of Visualized Experiments 59 doi:10.3791/3686