Protein-protein interactions monitored in mammalian cells via complementation of beta -lactamase enzyme fragments. Academic Article uri icon

Overview

abstract

  • We have defined inactive alpha and omega fragments of beta-lactamase that can complement to form a functional enzyme in both bacteria and mammalian cells, serving as a readout for the interaction of proteins fused to the fragments. Critical to this advance was the identification of a tripeptide, Asn-Gly-Arg, which when juxtaposed at the carboxyl terminus of the alpha fragment increased complemented enzyme activity by up to 4 orders of magnitude. beta-Lactamase is well suited to monitoring constitutive and inducible protein interactions because it is small (29 kDa), monomeric, and assayable with a fluorescent cell-permeable substrate. The negligible background, the magnitude of induced signal caused by enzymatic amplification, and detection of signal within minutes are unparalleled in mammalian protein interaction detection systems published to date.

publication date

  • March 19, 2002

Research

keywords

  • Escherichia coli
  • Genetic Complementation Test
  • Peptides
  • beta-Lactamases

Identity

PubMed Central ID

  • PMC122547

Scopus Document Identifier

  • 0037133629

Digital Object Identifier (DOI)

  • 10.1073/pnas.062043699

PubMed ID

  • 11904411

Additional Document Info

volume

  • 99

issue

  • 6