Abstract
The conformation of the pressure-dissociated monomer of Arc repressor was characterized by 1H NMR spectroscopy. The NMR spectra of the monomer under pressure (up to 5.0 kbar; 1 bar = 100 kPa) are typical of a molten globule and they are considerably different from those of the native dimer and thermally denatured monomer. The two-dimensional nuclear Overhauser effect spectra suggest that the pressure-induced molten globule retains some secondary structure. The presence of nuclear Overhauser effects in the beta-sheet region in the dissociated state suggests that the intermonomer beta-sheet (residues 8-14) in the native dimer is replaced by an intramonomer beta-sheet. Changes in one-dimensional and two-dimensional NMR spectra prior to pressure dissociation were found and suggest the existence of a "predissociated" state.
Full text
PDF




Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Baum J., Dobson C. M., Evans P. A., Hanley C. Characterization of a partly folded protein by NMR methods: studies on the molten globule state of guinea pig alpha-lactalbumin. Biochemistry. 1989 Jan 10;28(1):7–13. doi: 10.1021/bi00427a002. [DOI] [PubMed] [Google Scholar]
- Breg J. N., Boelens R., George A. V., Kaptein R. Sequence-specific 1H NMR assignment and secondary structure of the Arc repressor of bacteriophage P22, as determined by two-dimensional 1H NMR spectroscopy. Biochemistry. 1989 Dec 12;28(25):9826–9833. doi: 10.1021/bi00451a042. [DOI] [PubMed] [Google Scholar]
- Breg J. N., van Opheusden J. H., Burgering M. J., Boelens R., Kaptein R. Structure of Arc repressor in solution: evidence for a family of beta-sheet DNA-binding proteins. Nature. 1990 Aug 9;346(6284):586–589. doi: 10.1038/346586a0. [DOI] [PubMed] [Google Scholar]
- Creighton T. E. Protein folding. Biochem J. 1990 Aug 15;270(1):1–16. doi: 10.1042/bj2700001. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Creighton T. E. Toward a better understanding of protein folding pathways. Proc Natl Acad Sci U S A. 1988 Jul;85(14):5082–5086. doi: 10.1073/pnas.85.14.5082. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dill K. A., Shortle D. Denatured states of proteins. Annu Rev Biochem. 1991;60:795–825. doi: 10.1146/annurev.bi.60.070191.004051. [DOI] [PubMed] [Google Scholar]
- Heremans K. High pressure effects on proteins and other biomolecules. Annu Rev Biophys Bioeng. 1982;11:1–21. doi: 10.1146/annurev.bb.11.060182.000245. [DOI] [PubMed] [Google Scholar]
- Jaenicke R. Folding and association of proteins. Prog Biophys Mol Biol. 1987;49(2-3):117–237. doi: 10.1016/0079-6107(87)90011-3. [DOI] [PubMed] [Google Scholar]
- Kim P. S., Baldwin R. L. Intermediates in the folding reactions of small proteins. Annu Rev Biochem. 1990;59:631–660. doi: 10.1146/annurev.bi.59.070190.003215. [DOI] [PubMed] [Google Scholar]
- Ohgushi M., Wada A. 'Molten-globule state': a compact form of globular proteins with mobile side-chains. FEBS Lett. 1983 Nov 28;164(1):21–24. doi: 10.1016/0014-5793(83)80010-6. [DOI] [PubMed] [Google Scholar]
- Roder H. Structural characterization of protein folding intermediates by proton magnetic resonance and hydrogen exchange. Methods Enzymol. 1989;176:446–473. doi: 10.1016/0076-6879(89)76024-9. [DOI] [PubMed] [Google Scholar]
- Samarasinghe S. D., Campbell D. M., Jonas A., Jonas J. High-resolution NMR study of the pressure-induced unfolding of lysozyme. Biochemistry. 1992 Sep 1;31(34):7773–7778. doi: 10.1021/bi00149a005. [DOI] [PubMed] [Google Scholar]
- Silva J. L., Silveira C. F., Correia Júnior A., Pontes L. Dissociation of a native dimer to a molten globule monomer. Effects of pressure and dilution on the association equilibrium of arc repressor. J Mol Biol. 1992 Jan 20;223(2):545–555. doi: 10.1016/0022-2836(92)90669-b. [DOI] [PubMed] [Google Scholar]
- Vershon A. K., Bowie J. U., Karplus T. M., Sauer R. T. Isolation and analysis of arc repressor mutants: evidence for an unusual mechanism of DNA binding. Proteins. 1986 Dec;1(4):302–311. doi: 10.1002/prot.340010404. [DOI] [PubMed] [Google Scholar]
- Vershon A. K., Youderian P., Susskind M. M., Sauer R. T. The bacteriophage P22 arc and mnt repressors. Overproduction, purification, and properties. J Biol Chem. 1985 Oct 5;260(22):12124–12129. [PubMed] [Google Scholar]
- Weber G., Drickamer H. G. The effect of high pressure upon proteins and other biomolecules. Q Rev Biophys. 1983 Feb;16(1):89–112. doi: 10.1017/s0033583500004935. [DOI] [PubMed] [Google Scholar]
- Wishart D. S., Sykes B. D., Richards F. M. The chemical shift index: a fast and simple method for the assignment of protein secondary structure through NMR spectroscopy. Biochemistry. 1992 Feb 18;31(6):1647–1651. doi: 10.1021/bi00121a010. [DOI] [PubMed] [Google Scholar]
- Zagorski M. G., Bowie J. U., Vershon A. K., Sauer R. T., Patel D. J. NMR studies of Arc repressor mutants: proton assignments, secondary structure, and long-range contacts for the thermostable proline-8----leucine variant of Arc. Biochemistry. 1989 Dec 12;28(25):9813–9825. doi: 10.1021/bi00451a041. [DOI] [PubMed] [Google Scholar]