Stable Substructures in Proteins and How to Find Them Using Single-Molecule Force Spectroscopy

Katarzyna Tych, Gabriel Žoldák*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

1 Citation (Scopus)

Abstract

Three-dimensional structures of proteins are a source of fascination for scientists, due to the beauty of their sequence-encoded architectures and their highly diverse range of functions. These functions include acting as powerful catalysts, signal receptors, and versatile molecular motors as well as being building blocks for macroscopic structures, thus defining the shape of multicellular organisms. How protein structure is organized and assembled at the sub-nanometer scale is of great current interest. Specifically, the discovery of stable substructures and supersecondary structures has inspired research into their potential use in rationally engineered proteins with tailor-made properties. Here, we show how the search for stable substructures in large proteins can benefit from recent advances in single-molecule force spectroscopy using highly sensitive dual-beam optical tweezers. Our chapter provides a step-by-step description of the experimental workflow for (1) preparing proteins for mechanical interrogation, (2) interpreting the data, and (3) avoiding the most commonly occurring mistakes.

Original languageEnglish
Title of host publicationProtein Supersecondary Structures
Subtitle of host publicationMethods and Protocols
EditorsAlexander E. Kister
Place of PublicationNew York
PublisherHumana Press
Pages263-282
Number of pages20
ISBN (Electronic)978-1-4939-9161-7
ISBN (Print)978-1-4939-9160-0
DOIs
Publication statusPublished - 2019
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1958
ISSN (Print)1064-3745

Keywords

  • Amino Acid Motifs
  • Catalysis
  • Molecular Biology/methods
  • Molecular Conformation
  • Protein Engineering/methods
  • Proteins/chemistry

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