Site-specific biotinylation of purified proteins using BirA
Enzymatic biotinylation with E. coli biotin ligase (BirA) is highly specific in covalently attaching biotin to the 15 amino acid AviTag peptide, giving a homogeneous product with high yield. AviTag can conveniently be added genetically at the N-terminus, C-terminus, or …
Expression and purification of E. coli BirA biotin ligase for ... - PubMed
BirA is the Escherichia coli biotin ligase that site-specifically biotinylates a lysine side chain within a 15-amino acid acceptor peptide (also known as Avi-tag). As a complementary approach to in vivo biotinylation of Avi-tag-bearing proteins, we developed a protocol for producing recombinant BirA ligase for in vitro biotinylation.
Expression and purification of E. coli BirA biotin ligase for in …
BirA is the Escherichia coli biotin ligase that site-specifically biotinylates a lysine side chain within a 15-amino acid acceptor peptide (also known as Avi-tag). As a complementary approach to in vivo biotinylation of Avi-tag-bearing proteins, we developed a protocol for producing recombinant BirA ligase for in vitro biotinylation.
A proximity-dependent biotinylation map of a human cell | Nature
Jun 2, 2021 · In BioID, a mutant Escherichia coli biotin ligase (BirA*, R118G) is fused to a ‘bait’ polypeptide of interest, and the resulting fusion protein expressed in cultured cells or organisms 9.
Towards improving proximity labeling by the biotin ligase BirA
We found that relative to wild-type BirA, the Arg118Lys substitution both slightly reduced biotin affinity and increased the release of reactive biotinyl-5′-AMP. BioID using a BirA (Arg118Lys)-Lamin A fusion enabled identification of PCNA as a lamina-proximal protein in HEK293T cells, a finding that was validated by immunofluorescence microscopy.
A bacterial display system for effective selection of protein-biotin …
Mar 11, 2019 · Biotinylation creates a sensitive and specific tag for purification and detection of target proteins. The E. coli protein-biotin ligase BirA biotinylates a lysine within a synthetic biotin...
We expressed the bacterial BirA biotin ligase in mammalian cells and demonstrated very efficient biotinylation of a hematopoietic transcription factor bearing a small (23-aa) artificial peptide tag.
RNA–protein interaction detection in living cells - Nature
Feb 5, 2018 · Here we describe RNA–protein interaction detection (RaPID), which uses proximity-dependent protein labeling, based on the BirA* biotin ligase, to rapidly identify the proteins that bind RNA...
Expression and purification of E. coli BirA biotin ligase for in …
Mar 1, 2012 · BirA is the Escherichia coli biotin ligase that site-specifically biotinylates a lysine side chain within a 15-amino acid acceptor peptide (also known as Avi-tag). As a complementary approach to in vivo biotinylation of Avi-tag-bearing proteins, we developed a protocol for producing recombinant BirA ligase for in vitro biotinylation.
AirID, a novel proximity biotinylation enzyme, for analysis of …
May 11, 2020 · Here, we demonstrate a novel BirA enzyme, AirID (ancestral BirA for proximity-dependent biotin identification), which was designed de novo using an ancestral enzyme reconstruction algorithm and metagenome data.