New partners of acyl carrier protein detected in Escherichia coli by tandem affinity purification.
We report the first use of tandem affinity purification (TAP) in a prokaryote to purify native protein complexes, and demonstrate its reliability and power. We purified the acyl carrier protein (ACP) of Escherichia coli, a protein involved in a myriad of metabolic pathways. Besides the identification of several known partners of ACP, we rediscovered ACP/MukB and ACP/IscS interactions already detected but previously disregarded as due to contamination. Here, we demonstrate the specificity of these interactions and characterize them. This suggests that ACP is involved in additional previously unsuspected pathways. Furthermore, this study shows how the TAP method can be simply used in prokaryotes such as E. coli to identify new partners in protein-protein interactions under physiological conditions and thereby uncover novel protein functions.
Gully
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D
LISM, CNRS, 31 chemin Joseph Aiguier, 13402, Marseille, France.
Moinier
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D
Loiseau
L
L
Bouveret
E
E
D000213:Acyl Carrier Protein / Q000302:isolation & purification* / Q000378:metabolism*; D000345:Affinity Labels; D001426:Bacterial Proteins / Q000378:metabolism; D013437:Carbon-Sulfur Lyases / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone; D004220:Disulfides; D029968:Escherichia coli Proteins / Q000302:isolation & purification / Q000378:metabolism*; D008722:Methods; D008863:Microspheres; D011485:Protein Binding; D012694:Serine
D000213:Acyl Carrier Protein; D000345:Affinity Labels; D001426:Bacterial Proteins; D002868:Chromosomal Proteins, Non-Histone; D004220:Disulfides; D029968:Escherichia coli Proteins; C067035:MukB protein, E coli; D012694:Serine; D013437:Carbon-Sulfur Lyases; C080546:cysteine desulfurase