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Title: An on-pathway intermediate in the folding of a PDZ domain
Authors: Ivarsson, Ylva
Travaglini-Allocatelli, Carlo
Jemth, Per
Malatesta, Francesco
Brunori, Maurizio
Gianni, Stefano # ×
Issue Date: Mar-2007
Publisher: Amer soc biochemistry molecular biology inc
Series Title: Journal of biological chemistry vol:282 issue:12 pages:8568-8572
Abstract: The folding pathways of some proteins include the population of partially structured species en route to the native state. Identification and characterization of these folding intermediates are particularly difficult as they are often only transiently populated and play different mechanistic roles, being either on-pathway productive species or off-pathway kinetic traps. To define the role of folding intermediates, a quantitative analysis of the folding and unfolding rate constants over a wide range of denaturant concentration is often required. Such a task is further complicated by the reversible nature of the folding reaction, which implies the observed kinetics to be governed by a complex combination of different microscopic rate constants. Here, we tackled this problem by measuring directly the folding rate constant under highly denaturing conditions, namely by inducing the folding of a PDZ domain through a quasi-irreversible binding reaction with a specific peptide. In analogy with previous works based on hydrogen exchange experiments, we present evidence that the folding pathway of the PDZ domain involves the formation of an obligatory on-pathway intermediate. The results presented exemplify a novel type of kinetic test to detect an on-pathway folding intermediate.
ISSN: 0021-9258
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Molecular Genetics Section (-)
× corresponding author
# (joint) last author

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