Energetic landscape of alpha-lytic protease optimizes longevity through kinetic stability.
During the evolution of proteins the pressure to optimize biological activity is moderated by a need for efficient folding. For most proteins, this is accomplished through spontaneous folding to a thermodynamically stable and active native state. However, in the extracellular bacterial alpha-lytic protease (alphaLP) these two processes have become decoupled. The native state of alphaLP is thermodynamically unstable, and when denatured, requires millennia (t1/2 approximately 1,800 years) to refold. Folding is made possible by an attached folding catalyst, the pro-region, which is degraded on completion of folding, leaving alphaLP trapped in its native state by a large kinetic unfolding barrier (t1/2 approximately 1.2 years). alphaLP faces two very different folding landscapes: one in the presence of the pro-region controlling folding, and one in its absence restricting unfolding. Here we demonstrate that this separation of folding and unfolding pathways has removed constraints placed on the folding of thermodynamically stable proteins, and allowed the evolution of a native state having markedly reduced dynamic fluctuations. This, in turn, has led to a significant extension of the functional lifetime of alphaLP by the optimal suppression of proteolytic sensitivity.
Jaswal
Sheila S
SS
Department of Biochemistry and Biophysics, University of California at San Francisco, 94143-0448, USA.
Sohl
Julie L
JL
Davis
Jonathan H
JH
Agard
David A
DA
D000818:Animals; D004795:Enzyme Stability; D004926:Escherichia coli; D007700:Kinetics; D008958:Models, Molecular; D010843:Pichia; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry; D012697:Serine Endopeptidases / Q000737:chemistry* / Q000378:metabolism; D013816:Thermodynamics; D014867:Water / Q000737:chemistry
D011994:Recombinant Proteins; D014867:Water; D012697:Serine Endopeptidases; C021826:myxobacter alpha-lytic proteinase