Scripps VIVO scripps research logo

  • Index
  • Log in
  • Home
  • People
  • Organizations
  • Research
  • Events
Search form

Thermodynamics of a designed protein catenane

Academic Article
uri icon
  • Overview
  • Research
  • Identity
  • Additional Document Info
  • View All
scroll to property group menus

Overview

related to degree

  • Blankenship, John, Ph.D. in Macromolecular and Cellular Structure and Chemistry, Scripps Research 1997 - 2003

authors

  • Blankenship, John
  • Dawson, Philip

publication date

  • March 2003

journal

  • Journal of Molecular Biology  Journal

abstract

  • Topological linking of proteins is a new approach for stabilizing and controlling the oligomerization state of proteins that fold in an interwined manner. The recent design of a backbone cyclized protein catenane based on the p53tet domain suggested that topological cross-linking provided increased stability against thermal and chemical denaturation. However, the tetrameric structure complicated detailed biophysical analysis of this protein. Here, we describe the design, synthesis and thermodynamic characterization of a protein catenane based on a dimeric mutant of the p53tet domain (M340E/L344K). The formation of the catenane proceeded efficiently, and the overall structure and oligomerization of the domain was not affected by the formation of the topological link. Unfolding and refolding of the catenane was consistent with a two-state process. The topological link stabilized the dimer against thermal and chemical denaturation considerably, raising the apparent melting temperature by 59 degrees C and the midpoint of denaturation by 4.5M GuHCl at a concentration of 50 microM. The formation of the topological link increased the resistance of the dimer to proteolysis. However, the m value decreased by 1.7kcalmol(-1)M(-1), suggesting a decrease in accessible surface area in the unfolded state. This implies that the stabilization from the topological link is largely due to a destabilization of the unfolded state, similar to other cross-links in proteins. Topological linking therefore provides a powerful and orthogonal tool for the stabilization of peptide and protein oligomers.

subject areas

  • Anthracenes
  • Chromatography, High Pressure Liquid
  • Circular Dichroism
  • Cross-Linking Reagents
  • Dimerization
  • Disulfides
  • Guanidine
  • Humans
  • Mass Spectrometry
  • Models, Molecular
  • Peptide Fragments
  • Peptide Hydrolases
  • Polycyclic Compounds
  • Protein Conformation
  • Protein Denaturation
  • Protein Engineering
  • Protein Folding
  • Spectrometry, Fluorescence
  • Thermodynamics
  • Tumor Suppressor Protein p53
  • Ultracentrifugation
scroll to property group menus

Research

keywords

  • chemical ligation
  • protein design
  • protein stability
  • thermodynamics
  • topological chemistry
scroll to property group menus

Identity

International Standard Serial Number (ISSN)

  • 0022-2836

Digital Object Identifier (DOI)

  • 10.1016/s0022-2836(03)00115-3

PubMed ID

  • 12628256
scroll to property group menus

Additional Document Info

start page

  • 537

end page

  • 548

volume

  • 327

issue

  • 2

©2021 The Scripps Research Institute | Terms of Use | Powered by VIVO

  • About
  • Contact Us
  • Support