A simple method for probing the mechanical unfolding pathway of proteins in detail

• DOI: 10.1073/pnas.192351899
• Publication Date: 2002-09-17T16:40:55Z
• Authors: Robert B. Best; Susan B. Fowler; José L. Toca-Herrera; Jane Clarke
• Keywords: Barnase; Protein Engineering; Folding (DSP implementation)

Atomic force microscopy is an exciting new single-molecule technique to add the toolbox of protein (un)folding methods. However, detailed analysis unfolding proteins on application has, date, relied molecular dynamics simulations or a qualitative interpretation mutant data. Here we describe how engineering Φ value can be adapted characterize transition states for mechanical proteins. Single-molecule studies also have advantage over bulk experiments, in that partial values arising from structure state clearly distinguished those averaged alternate pathways. We show rate constants derived standard way by using Monte Carlo are not reliable because errors involved. it possible circumvent these problems, providing mechanism changed mutation, either modification procedure comparing and wild-type data directly. The applicability method tested simulated sets experimental mutants titin I27.