Nature 356, 263 - 265 (19 March 1992); doi:10.1038/356263a0

A protein-folding reaction under kinetic control

David Baker, Julie L. Sohl & David A. Agard

Howard Hughes Medical Institute, The Department of Biochemistry and the Biophysics Graduate Group, University of California at San Francisco, San Francisco, California 94143-0448, USA

SYNTHESIS of -lytic protease is as a precursor containing a 166 amino-acid pro region₁ transiently required for the correct folding of the protease domain^2–4. By omitting the pro region in an in vitrorefolding reaction we trapped an inactive, but folding competent state (I) having an expanded radius yet native-like secondary structure. The I state is stable for weeks at physiological pH in the absence of denaturant, but rapidly folds to the active, native state on addition of the pro region as a separate polypeptide chain. The mechanism of action of the pro region is distinct from that of the chaperonins^5,6: rather than reducing the rate of off-pathway reactions, the pro region accelerates the rate-limiting step on the folding pathway by more than 10⁷. Because both the I and native states are stable under identical conditions with no detectable interconversion, the folding of -lytic protease must be under kinetic and not thermodynamic control.

References

1.	Silen, J. L., McGrath, C. N., Smith, K. R. & Agard, D. A. Gene 69, 237−244 (1988). | Article | PubMed | ISI | ChemPort |
2.	Silen, J. L., Frank, D., Fujishige, A., Bone, R. & Agard, D. A. J. Bact. 171, 1320−1325 (1989). | PubMed | ChemPort |
3.	Silen, J. L. & Agard, D. A. Nature 341, 462−464 (1989). | Article | PubMed | ISI | ChemPort |
4.	Baker, D., Silen, J. L. & Agard, D. A. Proteins 12, 339−399 (1992). | Article | PubMed | ISI | ChemPort |
5.	Ellis, R. J. Nature 328, 378−379 (1987). | Article | PubMed | ISI | ChemPort |
6.	Goloubinoff, P., Christeller, J. T., Gatenby, A. A. & Lorimer, G. H. Nature 342, 884−889 (1989). | Article | PubMed | ISI | ChemPort |
7.	Goto, Y., Calciano, L. J. & Fink, A. L. Proc. natn. Acad. Sci. U.S.A. 87, 573−577 (1990). | ChemPort |
8.	Goto, Y., Takashi, N. & Fink, A. L. Biochemistry 29, 3480−3488 (1990). | Article | PubMed | ISI | ChemPort |
9.	Dolgikh, D. A. et al. FEBS Lett. 136, 311−315 (1981). | Article | PubMed | ISI | ChemPort |
10.	Kuwajima, K. Proteins 6, 87−103 (1989). | Article | PubMed | ISI | ChemPort |
11.	Kim, P. & Baldwin, R. L. A. Rev. Biochem. 59, 631−660 (1990). | Article | ISI | ChemPort |
12.	Dill, K. A. Biochemistry 29, 133−155 (1989).
13.	Kettner, C. A., Bone, R., Agard, D. A. & Bachovchin, W. B. Biochemistry 27, 7682−7688 (1988). | Article | PubMed | ChemPort |
14.	Fujinaga, M., Delbaere, L. T. J., Brayer, G. D. & James, M. N. J. molec. Biol. 183, 479−502 (1985). | PubMed |
15.	Kundrot, C. E. & Richards, F. M. Proteins Struct Funct. Genet. 3, 71−84 (1988). | PubMed |
16.	Roche, R. S. & Corbett, R. J. Biochemistry 23, 1888−1894 (1984). | PubMed | ISI | ChemPort |
17.	Ie Maire, M., Viel, A. & Moller, J. V. Analyt. Biochem. 177, 50−56 (1989). | PubMed |
18.	Chang, C. T., Wu, C. C. & Yang, J. T. Analyt Biochem. 91, 13−31 (1978). | PubMed | ISI | ChemPort |