Engineering proteins with enhanced mechanical stability by force-specific sequence motifs

Wenzhe Lu, Surendra S. Negi, Andres Oberhauser, Werner Braun

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Use of atomic force microscopy (AFM) has recently led to a better understanding of the molecular mechanisms of the unfolding process by mechanical forces; however, the rational design of novel proteins with specific mechanical strength remains challenging. We have approached this problem from a new perspective that generates linear physical-chemical properties (PCP) motifs from a limited AFM data set. Guided by our linear sequence analysis, we designed and analyzed four new mutants of the titin I1 domain with the goal of increasing the domain's mechanical strength. All four mutants could be cloned and expressed as soluble proteins. AFM data indicate that at least two of the mutants have increased molecular mechanical strength. This observation suggests that the PCP method is useful to graft sequences specific for high mechanical stability to weak proteins to increase their mechanical stability, and represents an additional tool in the design of novel proteins besides steered molecular dynamics calculations, coarse grained simulations, and φ{symbol}-value analysis of the transition state.

Original languageEnglish (US)
Pages (from-to)1308-1315
Number of pages8
JournalProteins: Structure, Function and Bioinformatics
Volume80
Issue number5
DOIs
StatePublished - May 2012

Fingerprint

Protein Engineering
Mechanical stability
Atomic Force Microscopy
Strength of materials
Atomic force microscopy
Chemical properties
Proteins
Mechanical Phenomena
Connectin
Value engineering
Molecular Dynamics Simulation
Grafts
Sequence Analysis
Molecular dynamics
Transplants

Keywords

  • Atomic force microscopy
  • CD spectroscopy
  • Molecular dynamics calculations
  • Physical-chemical property motifs
  • Protein design
  • Titin

ASJC Scopus subject areas

  • Biochemistry
  • Structural Biology
  • Molecular Biology

Cite this

Engineering proteins with enhanced mechanical stability by force-specific sequence motifs. / Lu, Wenzhe; Negi, Surendra S.; Oberhauser, Andres; Braun, Werner.

In: Proteins: Structure, Function and Bioinformatics, Vol. 80, No. 5, 05.2012, p. 1308-1315.

Research output: Contribution to journalArticle

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