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. 2000 Jun;9(6):1254–1261. doi: 10.1110/ps.9.6.1254

Assisted folding of D-glyceraldehyde-3-phosphate dehydrogenase by trigger factor.

G C Huang 1, Z Y Li 1, J M Zhou 1, G Fischer 1
PMCID: PMC2144665  PMID: 10892818

Abstract

The Escherichia coli trigger factor is a peptidyl-prolyl cis-trans isomerase that catalyzes proline-limited protein folding extremely well. Here, refolding of D-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in the presence of trigger factor was investigated. The regain of activity of GAPDH was markedly increased by trigger factor after either long- or short-term denaturation, and detectable aggregation of GAPDH intermediates was prevented. In both cases, time courses of refolding of GAPDH were decelerated by trigger factor. The reactivation yield of GAPDH showed a slow down-turn when molar ratios of trigger factor to GAPDH were above 5, due to tight binding between trigger factor and GAPDH intermediates. Such inactive bound GAPDH could be partially rescued from trigger factor by addition of reduced alphaLA as competitor, by further diluting the refolding mixture, or by disrupting hydrophobic interactions in the complexes. A model for trigger factor assisted refolding of GAPDH is proposed. We also suggest that assisted refolding of GAPDH is due mainly to the chaperone function of trigger factor.

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Selected References

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  1. Cai H., Wang C. C., Tsou C. L. Chaperone-like activity of protein disulfide isomerase in the refolding of a protein with no disulfide bonds. J Biol Chem. 1994 Oct 7;269(40):24550–24552. [PubMed] [Google Scholar]
  2. Crooke E., Guthrie B., Lecker S., Lill R., Wickner W. ProOmpA is stabilized for membrane translocation by either purified E. coli trigger factor or canine signal recognition particle. Cell. 1988 Sep 23;54(7):1003–1011. doi: 10.1016/0092-8674(88)90115-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Crooke E., Wickner W. Trigger factor: a soluble protein that folds pro-OmpA into a membrane-assembly-competent form. Proc Natl Acad Sci U S A. 1987 Aug;84(15):5216–5220. doi: 10.1073/pnas.84.15.5216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Deuerling E., Schulze-Specking A., Tomoyasu T., Mogk A., Bukau B. Trigger factor and DnaK cooperate in folding of newly synthesized proteins. Nature. 1999 Aug 12;400(6745):693–696. doi: 10.1038/23301. [DOI] [PubMed] [Google Scholar]
  5. Ewbank J. J., Creighton T. E. Pathway of disulfide-coupled unfolding and refolding of bovine alpha-lactalbumin. Biochemistry. 1993 Apr 13;32(14):3677–3693. doi: 10.1021/bi00065a022. [DOI] [PubMed] [Google Scholar]
  6. Ewbank J. J., Creighton T. E. Structural characterization of the disulfide folding intermediates of bovine alpha-lactalbumin. Biochemistry. 1993 Apr 13;32(14):3694–3707. doi: 10.1021/bi00065a023. [DOI] [PubMed] [Google Scholar]
  7. Fischer G., Bang H., Mech C. Nachweis einer Enzymkatalyse für die cis-trans-Isomerisierung der Peptidbindung in prolinhaltigen Peptiden. Biomed Biochim Acta. 1984;43(10):1101–1111. [PubMed] [Google Scholar]
  8. Freskgård P. O., Bergenhem N., Jonsson B. H., Svensson M., Carlsson U. Isomerase and chaperone activity of prolyl isomerase in the folding of carbonic anhydrase. Science. 1992 Oct 16;258(5081):466–468. doi: 10.1126/science.1357751. [DOI] [PubMed] [Google Scholar]
  9. Gill S. C., von Hippel P. H. Calculation of protein extinction coefficients from amino acid sequence data. Anal Biochem. 1989 Nov 1;182(2):319–326. doi: 10.1016/0003-2697(89)90602-7. [DOI] [PubMed] [Google Scholar]
  10. Guthrie B., Wickner W. Trigger factor depletion or overproduction causes defective cell division but does not block protein export. J Bacteriol. 1990 Oct;172(10):5555–5562. doi: 10.1128/jb.172.10.5555-5562.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Göthel S. F., Scholz C., Schmid F. X., Marahiel M. A. Cyclophilin and trigger factor from Bacillus subtilis catalyze in vitro protein folding and are necessary for viability under starvation conditions. Biochemistry. 1998 Sep 22;37(38):13392–13399. doi: 10.1021/bi981253w. [DOI] [PubMed] [Google Scholar]
  12. Hesterkamp T., Hauser S., Lütcke H., Bukau B. Escherichia coli trigger factor is a prolyl isomerase that associates with nascent polypeptide chains. Proc Natl Acad Sci U S A. 1996 Apr 30;93(9):4437–4441. doi: 10.1073/pnas.93.9.4437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kandror O., Sherman M., Moerschell R., Goldberg A. L. Trigger factor associates with GroEL in vivo and promotes its binding to certain polypeptides. J Biol Chem. 1997 Jan 17;272(3):1730–1734. doi: 10.1074/jbc.272.3.1730. [DOI] [PubMed] [Google Scholar]
  14. Kandror O., Sherman M., Rhode M., Goldberg A. L. Trigger factor is involved in GroEL-dependent protein degradation in Escherichia coli and promotes binding of GroEL to unfolded proteins. EMBO J. 1995 Dec 1;14(23):6021–6027. doi: 10.1002/j.1460-2075.1995.tb00290.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kern G., Kern D., Schmid F. X., Fischer G. A kinetic analysis of the folding of human carbonic anhydrase II and its catalysis by cyclophilin. J Biol Chem. 1995 Jan 13;270(2):740–745. doi: 10.1074/jbc.270.2.740. [DOI] [PubMed] [Google Scholar]
  16. Kofron J. L., Kuzmic P., Kishore V., Colón-Bonilla E., Rich D. H. Determination of kinetic constants for peptidyl prolyl cis-trans isomerases by an improved spectrophotometric assay. Biochemistry. 1991 Jun 25;30(25):6127–6134. doi: 10.1021/bi00239a007. [DOI] [PubMed] [Google Scholar]
  17. Lang K., Schmid F. X., Fischer G. Catalysis of protein folding by prolyl isomerase. Nature. 1987 Sep 17;329(6136):268–270. doi: 10.1038/329268a0. [DOI] [PubMed] [Google Scholar]
  18. Li X. L., Lei X. D., Cai H., Li J., Yang S. L., Wang C. C., Tsou C. L. Binding of a burst-phase intermediate formed in the folding of denatured D-glyceraldehyde-3-phosphate dehydrogenase by chaperonin 60 and 8-anilino-1-naphthalenesulphonic acid. Biochem J. 1998 Apr 15;331(Pt 2):505–511. doi: 10.1042/bj3310505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Liang S. J., Lin Y. Z., Zhou J. M., Tsou C. L., Wu P. Q., Zhou Z. K. Dissociation and aggregation of D-glyceraldehyde-3-phosphate dehydrogenase during denaturation by guanidine hydrochloride. Biochim Biophys Acta. 1990 Apr 19;1038(2):240–246. doi: 10.1016/0167-4838(90)90211-w. [DOI] [PubMed] [Google Scholar]
  20. Lin L. N., Hasumi H., Brandts J. F. Catalysis of proline isomerization during protein-folding reactions. Biochim Biophys Acta. 1988 Oct 12;956(3):256–266. doi: 10.1016/0167-4838(88)90142-2. [DOI] [PubMed] [Google Scholar]
  21. Lyon W. R., Gibson C. M., Caparon M. G. A role for trigger factor and an rgg-like regulator in the transcription, secretion and processing of the cysteine proteinase of Streptococcus pyogenes. EMBO J. 1998 Nov 2;17(21):6263–6275. doi: 10.1093/emboj/17.21.6263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Scheek R. M., Slater E. C. Glyceraldehyde-3-phosphate dehydrogenase from rabbit muscle. Methods Enzymol. 1982;89(Pt 500):305–309. doi: 10.1016/s0076-6879(82)89055-1. [DOI] [PubMed] [Google Scholar]
  23. Schmid F. X., Mayr L. M., Mücke M., Schönbrunner E. R. Prolyl isomerases: role in protein folding. Adv Protein Chem. 1993;44:25–66. doi: 10.1016/s0065-3233(08)60563-x. [DOI] [PubMed] [Google Scholar]
  24. Scholz C., Stoller G., Zarnt T., Fischer G., Schmid F. X. Cooperation of enzymatic and chaperone functions of trigger factor in the catalysis of protein folding. EMBO J. 1997 Jan 2;16(1):54–58. doi: 10.1093/emboj/16.1.54. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Song J., Quan H., Wang C. Dependence of the anti-chaperone activity of protein disulphide isomerase on its chaperone activity. Biochem J. 1997 Dec 15;328(Pt 3):841–846. doi: 10.1042/bj3280841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Stoller G., Rücknagel K. P., Nierhaus K. H., Schmid F. X., Fischer G., Rahfeld J. U. A ribosome-associated peptidyl-prolyl cis/trans isomerase identified as the trigger factor. EMBO J. 1995 Oct 16;14(20):4939–4948. doi: 10.1002/j.1460-2075.1995.tb00177.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Teipel J. W., Koshland D. E., Jr Kinetic aspects of conformational changes in proteins. I. Rate of regain of enzyme activity from denatured proteins. Biochemistry. 1971 Mar 2;10(5):792–798. doi: 10.1021/bi00781a011. [DOI] [PubMed] [Google Scholar]
  28. Valent Q. A., Kendall D. A., High S., Kusters R., Oudega B., Luirink J. Early events in preprotein recognition in E. coli: interaction of SRP and trigger factor with nascent polypeptides. EMBO J. 1995 Nov 15;14(22):5494–5505. doi: 10.1002/j.1460-2075.1995.tb00236.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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