CPL - Chalmers Publication Library
| Utbildning | Forskning | Styrkeområden | Om Chalmers | In English In English Ej inloggad.

Probing the influence on folding behavior of structurally conserved core residues in P. aeruginosa apo-azurin.

Cecilia Engman ; Anders Sandberg ; Johan Leckner (Institutionen för kemi och biovetenskap, Molekylär bioteknik) ; B Göran Karlsson (Institutionen för kemi och biovetenskap, Molekylär bioteknik)
Protein Science Vol. 13 (2004), 10, p. 2706-2715.
[Artikel, refereegranskad vetenskaplig]

The effects on folding kinetics and equilibrium stability of core mutations in the apo-mutant C112S of azurin from Pseudomonas aeruginosa were studied. A number of conserved residues within the cupredoxin family were recognized by sequential alignment as constituting a common hydrophobic core: I7, F15, L33, W48, F110, L50, V95, and V31. Of these, I7, V31, L33, and L50 were mutated for the purpose of obtaining information on the transition state and a potential folding nucleus. In addition, residue V5 in the immediate vicinity of the common core, as well as T52, separate from the core, were mutated as controls. All mutants exhibited a nonlinear dependence of activation free energy of folding on denaturant concentration, although the refolding kinetics of the V31A/C112S mutant indicated that the V31A mutation destabilizes the transition state enough to allow folding via a parallel transition state ensemble. Phi-values could be calculated for three of the six mutants, V31A/C112S, L33A/C112S, and L50A/C112S, and the fractional values of 0.63, 0.33, and 0.50 (respectively) obtained at 0.5 M GdmCl suggest that these residues are important for stabilizing the transition state. Furthermore, a linear dependence of ln k(obs)(H2O) on DeltaG(U-N)(H2O) of the core mutations and the putative involvement of ground-state effects suggest the presence of native-like residual interactions in the denatured state that bias this ensemble toward a folding-competent state.

Nyckelord: Protein folding, cupredoxin



Denna post skapades 2006-08-29. Senast ändrad 2011-01-20.
CPL Pubid: 2871

 

Institutioner (Chalmers)

Institutionen för kemi (2001-2011)
Institutionen för kemi och biovetenskap, Molekylär bioteknik (2002-2004)

Ämnesområden

Industriell bioteknik

Chalmers infrastruktur