Format

Send to

Choose Destination
Protein Sci. 2004 Nov;13(11):3043-50. Epub 2004 Sep 30.

Promiscuous protein biotinylation by Escherichia coli biotin protein ligase.

Author information

1
Departments of Microbiology and Biochemistry, University of Illinois, Urbana, IL 61801, USA.

Abstract

Biotin protein ligases (BPLs) are enzymes of extraordinary specificity. BirA, the BPL of Escherichia coli biotinylates only a single cellular protein. We report a mutant BirA that attaches biotin to a large number of cellular proteins in vivo and to bovine serum albumin, chloramphenicol acetyltransferase, immunoglobin heavy and light chains, and RNAse A in vitro. The mutant BirA also self biotinylates in vivo and in vitro. The wild type BirA protein is much less active in these reactions. The biotinylation reaction is proximity-dependent in that a greater extent of biotinylation was seen when the mutant ligase was coupled to the acceptor proteins than when the acceptors were free in solution. This approach may permit facile detection and recovery of interacting proteins by existing avidin/streptavidin technology.

PMID:
15459338
PMCID:
PMC2286582
DOI:
10.1110/ps.04911804
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Wiley Icon for PubMed Central
Loading ...
Support Center