Asset Details
Do you wish to reserve the book?
Kinetic evidence for a two-stage mechanism of protein denaturation by guanidinium chloride
Hey, we have placed the reservation for you!
By the way, why not check out events that you can attend while you pick your title.
Oops! Something went wrong.
Looks like we were not able to place the reservation. Kindly try again later.
Are you sure you want to remove the book from the shelf?
Kinetic evidence for a two-stage mechanism of protein denaturation by guanidinium chloride
Do you wish to request the book?
Kinetic evidence for a two-stage mechanism of protein denaturation by guanidinium chloride
Please be aware that the book you have requested cannot be checked out. If you would like to checkout this book, you can reserve another copy
We have requested the book for you!
Your request is successful and it will be processed during the Library working hours. Please check the status of your request in My Requests.
Oops! Something went wrong.
Looks like we were not able to place your request. Kindly try again later.
Journal Article
Kinetic evidence for a two-stage mechanism of protein denaturation by guanidinium chloride
2014
Overview
Dry molten globular (DMG) intermediates, an expanded form of the native protein with a dry core, have been observed during denaturant-induced unfolding of many proteins. These observations are counterintuitive because traditional models of chemical denaturation rely on changes in solvent-accessible surface area, and there is no notable change in solvent-accessible surface area during the formation of the DMG. Here we show, using multisite fluorescence resonance energy transfer, far-UV CD, and kinetic thiol-labeling experiments, that the guanidinium chloride (GdmCl)-induced unfolding of RNase H also begins with the formation of the DMG. Population of the DMG occurs within the 5-ms dead time of our measurements. We observe that the size and/or population of the DMG is linearly dependent on [GdmCl], although not as strongly as the second and major step of unfolding, which is accompanied by core solvation and global unfolding. This rapid GdmCl-dependent population of the DMG indicates that GdmCl can interact with the protein before disrupting the hydrophobic core. These results imply that the effect of chemical denaturants cannot be interpreted solely as a disruption of the hydrophobic effect and strongly support recent computational studies, which hypothesize that chemical denaturants first interact directly with the protein surface before completely unfolding the protein in the second step (direct interaction mechanism).
Publisher
National Academy of Sciences,National Acad Sciences
Subject
/ Energy
/ Escherichia coli - enzymology
/ Globules
/ Kinetics
/ Nitrobenzoates - pharmacology
/ Protein Denaturation - drug effects
/ Protein Structure, Secondary
/ Protein Unfolding - drug effects
/ Proteins
/ Salt
/ Solvents
/ Sulfhydryl Compounds - metabolism
/ Sulfhydryl Compounds - pharmacology
/ Thiols
