Lanthanide-binding tags as versatile protein coexpression probes.

Published

Journal Article

Comprehensive proteomic analyses require new methodologies to accelerate the correlation of gene sequence with protein function. Key tools for such efforts include biophysical probes that integrate into the covalent architecture of proteins. Lanthanide-binding tags (LBTs) are expressible, multitasking fusion partners that are optimized to bind lanthanide ions and have several desirable attributes, which include long-lived luminescence, excellent X-ray scattering power for phase determination, and magnetic properties to facilitate NMR spectroscopic structure elucidation. Herein, we present peptide sequences with a 40-fold higher affinity for Tb(3+) ions and significantly brighter luminescence intensity compared with existing peptides. Incorporation of an LBT onto ubiquitin as a prototype fusion protein allows the use of powerful protein-visualization techniques, which include rapid luminescence detection of LBT-tagged proteins in SDS-PAGE gels, as well as determination of protein concentrations in complex mixtures. The LBT strategy is a new alternative for expressing fluorescent fusion proteins by routine molecular biological techniques.

Full Text

Duke Authors

Cited Authors

  • Franz, KJ; Nitz, M; Imperiali, B

Published Date

  • April 2003

Published In

Volume / Issue

  • 4 / 4

Start / End Page

  • 265 - 271

PubMed ID

  • 12672105

Pubmed Central ID

  • 12672105

Electronic International Standard Serial Number (EISSN)

  • 1439-7633

International Standard Serial Number (ISSN)

  • 1439-4227

Digital Object Identifier (DOI)

  • 10.1002/cbic.200390046

Language

  • eng