Promiscuous protein biotinylation by Escherichia coli biotin protein ligase.

  • Choi-Rhee E
  • Schulman H
  • Cronan J
  • 244


    Mendeley users who have this article in their library.
  • 59


    Citations of this article.


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.

Author-supplied keywords

  • acyl adenylate
  • acylating
  • biotin protein ligase
  • biotinylation
  • biotinylation of proteins has
  • bira
  • cal means
  • chemi-
  • protein amino groups
  • protein modification
  • routinely been done by
  • usually by modification of
  • with biotin-n-hydroxysuccinimide or similar

Get free article suggestions today

Mendeley saves you time finding and organizing research

Sign up here
Already have an account ?Sign in

Find this document


  • Eunjoo Choi-Rhee

  • Howard Schulman

  • John E Cronan

Cite this document

Choose a citation style from the tabs below

Save time finding and organizing research with Mendeley

Sign up for free