The largest protein observed to fold by two-state kinetic mechanism does not obey contact-order correlation

Journal of the American Chemical Society

Volumn 125, Issue 32, 2003, Pages 9606-9607

  Jones, Kathryn ^a   Wittung Stafshede, Pernilla ^a  

^a Tulane University   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

PROTEIN;

ARTICLE; BORRELIA BURGDORFERI; CHEMICAL REACTION KINETICS; CORRELATION ANALYSIS; PROTEIN ANALYSIS; PROTEIN DOMAIN; PROTEIN EXPRESSION; PROTEIN FOLDING; PROTEIN STRUCTURE; THERMODYNAMICS;

ANTIGENS, BACTERIAL; BACTERIAL PROTEINS; BORRELIA BURGDORFERI; CIRCULAR DICHROISM; KINETICS; LIPOPROTEINS; PROTEIN FOLDING; PROTEIN STRUCTURE, SECONDARY; PROTEIN STRUCTURE, TERTIARY; THERMODYNAMICS;

EID: 0041624356     PISSN: 00027863     EISSN: None     Source Type: Journal    
DOI: 10.1021/ja0358807     Document Type: Article

References (17)

1. Zhang, J. R.; Hardham, J. M.; Barbour, A. G.; Norris, S. J. Cell 1997, 89, 275-285.
2. Eicken, C.; Sharma, V.; Klabunde, T.; Lawrenz, M. B.; Hardham, J. M.; Norris, S. J.; Sacchettini, J. C. J. Biol. Chem. 2002, 277, 21691-6.
3. Jones, K.; Guidry, J.; Wittung-Stafshede, P. Biochem. Biophys. Res. Commun. 2001, 289, 389-94.
4. Plaxco, K. W.; Simons, K.; Ruczinski, I.; Baker, D. Biochemistry 2000, 39, 11177-83.
5. Pande, V. S. Proc. Natl. Acad. Sci. U.S.A. 2003, 100 (7), 3555-3556.
6. Recombinant VlsE was prepared as in ref 3. Equilibrium unfolding (10-100 μM VlsE) was performed at 20 °C, 5 mM phosphate, pH 7. Far-UV CD was measured on an OLIS spectrometer, tyrosine fluorescence on a Varian Eclipse. Guanidine hydrochloride (GuHCl) and urea were of highest purity. Data were analyzed in terms of two-state mechanisms.
7. Creighton, T. E., Ed. Protein Folding; Freeman & Co.: New York, 1992.
8. Ganesh, C.; Shah, A.; Swaminathan, C.; Surolia, A.; Varadarajan, R. Biochemistry 1997, 36, 5020-5028.
9. Feller, G.; d'Amico, D.; Gerday, C. Biochemistry 1999, 38, 4613-4619.
10. Lawrenz, M. B.; Hardham, J. M.; Owens, R. T.; Nowakowski, J.; Steere, A. C.; Wormser, G. P.; Norris, S. J. J. Clin. Microbiol. 1999, 37 (12), 3997-4004.
11. Stopped-flow experiments (20 °C, pH 7) were performed on a PiStar (Applied PhotoPhysics), monitoring fluorescence at 320 nm (ex 280 nm) and CD at 220 nm (2 mm cell, 1:5 mixing, final VlsE concentration 10 and 100 μM). Kinetics were monitored by fluorescence and CD for 17 GuHCl and 16 urea concentrations. In all cases, traces were best fit to single-exponential equations and CD and fluorescence data gave identical results. In Figure 2, the average rate constant at each condition is shown.
12. Apiyo, D.; Wittung-Stafshede, P. Protein Sci. 2002, 11, 1129-35.
13. http://depts.washington.edu/bakerpg/contact_order.
14. Wittung-Stafshede, P.; Lee, J. C.; Winkler, J. R.; Gray, H. B. Proc. Natl. Acad. Sci. U.S.A. 1999, 96, 6587-90.
15. Markarov, D. E.; Plaxco, K. W. Protein Sci. 2003, 12, 17-26.
16. Zagrovc, B.; Snow, C. D.; Khaliq, S.; Shirts, M. R.; Pande, V. S. J. Mol. Biol. 2002, 323, 153-164.
17. Robinson, C. R.; Sauer, R. T. Proc. Natl. Acad. Sci. U.S,A. 1998, 95, 5929-5934.