Mechanical unfolding of an ankyrin repeat protein.

Journal Academic Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't

Ankryin repeat proteins comprise tandem arrays of a 33-residue, predominantly alpha-helical motif that stacks roughly linearly to produce elongated and superhelical structures. They function as scaffolds mediating a diverse range of protein-protein interactions, and some have been proposed to play a role in mechanical signal transduction processes in the cell. Here we use atomic force microscopy and molecular-dynamics simulations to investigate the natural 7-ankyrin repeat protein gankyrin. We find that gankyrin unfolds under force via multiple distinct pathways. The reactions do not proceed in a cooperative manner, nor do they always involve fully stepwise unfolding of one repeat at a time. The peeling away of half an ankyrin repeat, or one or more ankyrin repeats, occurs at low forces; however, intermediate species are formed that are resistant to high forces, and the simulations indicate that in some instances they are stabilized by nonnative interactions. The unfolding of individual ankyrin repeats generates a refolding force, a feature that may be more easily detected in these proteins than in globular proteins because the refolding of a repeat involves a short contraction distance and incurs a low entropic cost. We discuss the origins of the differences between the force- and chemical-induced unfolding pathways of ankyrin repeat proteins, as well as the differences between the mechanics of natural occurring ankyrin repeat proteins and those of designed consensus ankyin repeat and globular proteins.

Full Text

Duke Authors

Cited Authors

  • Serquera, D; Lee, W; Settanni, G; Marszalek, PE; Paci, E; Itzhaki, LS

Published Date

  • April 7, 2010

Published In

Volume / Issue

  • 98 / 7

Start / End Page

  • 1294 - 1301

PubMed ID

  • 20371329

Electronic International Standard Serial Number (EISSN)

  • 1542-0086

Digital Object Identifier (DOI)

  • 10.1016/j.bpj.2009.12.4287

Language

  • eng

Citation Source

  • PubMed