Kinetic stability as a mechanism for protease longevity.
The folding of the extracellular serine protease, alpha-lytic protease (alphaLP; EC 3.4.21.12) reveals a novel mechanism for stability that appears to lead to a longer functional lifetime for the protease. For alphaLP, stability is based not on thermodynamics, but on kinetics. Whereas this has required the coevolution of a pro region to facilitate folding, the result has been the optimization of native-state properties independent of their consequences on thermodynamic stability. Structural and mutational data lead to a model for catalysis of folding in which the pro region binds to a conserved beta-hairpin in the alphaLP C-terminal domain, stabilizing the folding transition state and the native state. The pro region is then proteolytically degraded, leaving the active alphaLP trapped in a metastable conformation. This metastability appears to be a consequence of pressure to evolve properties of the native state, including a large, highly cooperative barrier to unfolding, and extreme rigidity, that reduce susceptibility to proteolytic degradation. In a test of survival under highly proteolytic conditions, homologous mammalian proteases that have not evolved kinetic stability are much more rapidly degraded than alphaLP. Kinetic stability as a means to longevity is likely to be a mechanism conserved among the majority of extracellular bacterial pro-proteases and may emerge as a general strategy for intracellular eukaryotic proteases subject to harsh conditions as well.
Cunningham
E L
EL
Graduate Group in Biophysics, Howard Hughes Medical Institute, Department of Biochemistry, University of California, San Francisco, CA 94143-0448, USA.
Jaswal
S S
SS
Sohl
J L
JL
Agard
D A
DA
D004795:Enzyme Stability; D007700:Kinetics; D017510:Protein Folding; D012697:Serine Endopeptidases / Q000737:chemistry* / Q000378:metabolism
D012697:Serine Endopeptidases; C021826:myxobacter alpha-lytic proteinase