Engineering of solvent-exposed loops in Escherichia coli β-galactosidase

Research output: Contribution to journalArticleResearchpeer-review

44 Citations (Scopus)

Abstract

The Escherichia coli β-galactosidase is a high molecular mass tetrameric enzyme extensively used as a molecular marker. Despite its proven utility as a partner in fusion proteins, previous attempts to generate insertional mutants rendered inactive or poorly active enzymes, hampering its further engineering for the construction of multifunctional enzymes. We have explored several solvent-exposed loops on the tetramer, namely those spanning residues 246-254, 271-287, 578-584, 770-773, and 793-806, as acceptor sites to accommodate functional protein segments on the surface of active β-galactosidase enzymes. An RGD-containing antigenic peptide positioned in these sites interacts efficiently with specific monoclonal antibodies as well as target integrins on the surface of mammalian cells. The resulting chimeric enzymes are soluble, stable, produced in high yields and enzymatically active. Moreover, the identified insertion sites could be appropriated for the design of promising β-galactosidase-based molecular sensors. Copyright (C) 1998 Federation of European Biochemical Societies.
Original languageEnglish
Pages (from-to)23-27
JournalFEBS Letters
Volume434
DOIs
Publication statusPublished - 28 Aug 1998

Keywords

  • Loop structure
  • Peptide display
  • Protein engineering
  • β-Galactosidase

Fingerprint

Dive into the research topics of 'Engineering of solvent-exposed loops in Escherichia coli β-galactosidase'. Together they form a unique fingerprint.

Cite this