Understanding and applications of Ser/Gly linkers in protein engineering

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

Abstract

Peptide linkers consisting of repeats of glycine and serine residues are commonly chosen by protein engineers to introduce flexible and hydrophilic spacers between protein domains. Given the popularity of these linkers, gaining a quantitative insight in their conformational behavior is important to understand the effect on functional properties of fusion proteins, including energy transfer efficiency in luminescent sensor proteins, intramolecular domain interactions and (multivalent) binding. In this chapter, we discuss how the conformational behavior of Ser/Gly linkers can be described using random coil models, and how measuring FRET as a function of linker length can be used to obtain empirical values for the stiffness of linkers containing different Ser-to-Gly ratios. Subsequently, we show how these models and the experimentally determined linker stiffness can be used to explain and predict the functional properties of multidomain proteins, providing useful rules-of-thumb and design tools for optimal linker engineering.

Original languageEnglish
Title of host publicationLinkers in Biomacromolecules
EditorsMaarten Merkx
PublisherAcademic Press Inc.
Pages1-22
Number of pages22
ISBN (Print)9780128208182
DOIs
Publication statusPublished - Jan 2021

Publication series

NameMethods in Enzymology
Volume647
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

  • FRET
  • Gaussian chain
  • Protein engineering
  • Protein switches
  • Ser/Gly linkers

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