Protein folding kinetics: barrier effects in chemical and thermal denaturation experiments

10 pages, 5 figures.-- PMID: 17419630 [PubMed].-- PMCID: PMC2527040.-- Author manuscript available in PMC: http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=17419630Printed version published on May 2, 2007.Recent experimental work on fast protein folding brings about an intriguing paradox. Microsecond-folding proteins are supposed to fold near or at the folding speed limit (downhill folding), but yet their folding behavior seems to comply with classical two-state analyses, which imply the crossing of high free energy barriers. However, close inspection of chemical and thermal denaturation kinetic experiments in fast-folding proteins reveals systematic deviations from two-state behavior. Using a simple one-dimensional free energy surface approach we find that such deviations are indeed diagnostic of marginal folding barriers. Furthermore, the quantitative analysis of available fast-kinetic data indicates that many microsecond-folding proteins fold downhill in native conditions. All of these proteins are then promising candidates for an atom-by-atom analysis of protein folding using nuclear magnetic resonance. We also find that the diffusion coefficient for protein folding is strongly temperature dependent, corresponding to an activation energy of ~1 kJ·mol-1 per protein residue. As a consequence, the folding speed limit at room temperature is about an order of magnitude slower than the ~ 1 μs estimates from high-temperature T-jump experiments. Our analysis is quantitatively consistent with the available thermodynamic and kinetic data on slow two-state folding proteins and provides a straightforward explanation for the apparent fast-folding paradox.This research has been supported by NIH grant GM066800-1 and NSF grant MCB-0317294.Peer reviewe

Evolution of Microstructure and Texture during Warm Rolling Of a Duplex Steel

The effect of warm rolling on the evolution of microstructure and texture in a duplex stainless steel (DSS) was investigated. For this purpose, a DSS steel was warm rolled up to 90 pct reduction in thickness at 498 K, 698 K, and 898 K (225 °C, 425 °C, and 625 °C). The microstructure with an alternate arrangement of deformed ferrite and austenite bands was observed after warm rolling; however, the microstructure after 90 pct warm rolling at 498 K and 898 K (225 °C and 625 °C) was more lamellar and uniform as compared to the rather fragmented and inhomogeneous structure observed after 90 pct warm rolling at 698 K (425 °C). The texture of ferrite in warm-rolled DSS was characterized by the presence of the RD (〈011〉//RD) and ND (〈111〉//ND) fibers. However, the texture of ferrite in DSS warm rolled at 698 K (425 °C) was distinctly different having much higher fraction of the RD-fiber components than that of the ND-fiber components. The texture and microstructural differences in ferrite in DSS warm rolled at different temperatures could be explained by the interaction of carbon atoms with dislocations. In contrast, the austenite in DSS warm rolled at different temperatures consistently showed pure metal- or copper-type deformation texture which was attributed to the increase in stacking fault energy at the warm-rolling temperatures. It was concluded that the evolution of microstructure and texture of the two constituent phases in DSS was greatly affected by the temperature of warm rolling, but not significantly by the presence of the other phas