TY - JOUR
T1 - Holo structure and steady state kinetics of the thiazolinyl imine reductases for siderophore biosynthesis
AU - Meneely, Kathleen M.
AU - Ronnebaum, Trey A.
AU - Riley, Andrew P.
AU - Prisinzano, Thomas E.
AU - Lamb, Audrey L.
N1 - Publisher Copyright:
© 2016 American Chemical Society.
PY - 2016/9/27
Y1 - 2016/9/27
N2 - Thiazolinyl imine reductases catalyze the NADPH-dependent reduction of a thiazoline to a thiazolidine, a required step in the formation of the siderophores yersiniabactin (Yersinia spp.) and pyochelin (Pseudomonas aeruginosa). These stand-alone nonribosomal peptide tailoring domains are structural homologues of sugar oxidoreductases. Two closed structures of the thiazolinyl imine reductase from Yersinia enterocolitica (Irp3) are presented here: an NADP+-bound structure to 1.45 Å resolution and a holo structure to 1.28 Å resolution with NADP+ and a substrate analogue bound. Michaelis-Menten kinetics were measured using the same substrate analogue and the homologue from P. aeruginosa, PchG. The data presented here support the hypothesis that tyrosine 128 is the likely general acid residue for catalysis and also highlight the phosphopantetheine tunnel for tethering of the substrate to the nonribosomal peptide synthetase module during assembly line biosynthesis of the siderophore.
AB - Thiazolinyl imine reductases catalyze the NADPH-dependent reduction of a thiazoline to a thiazolidine, a required step in the formation of the siderophores yersiniabactin (Yersinia spp.) and pyochelin (Pseudomonas aeruginosa). These stand-alone nonribosomal peptide tailoring domains are structural homologues of sugar oxidoreductases. Two closed structures of the thiazolinyl imine reductase from Yersinia enterocolitica (Irp3) are presented here: an NADP+-bound structure to 1.45 Å resolution and a holo structure to 1.28 Å resolution with NADP+ and a substrate analogue bound. Michaelis-Menten kinetics were measured using the same substrate analogue and the homologue from P. aeruginosa, PchG. The data presented here support the hypothesis that tyrosine 128 is the likely general acid residue for catalysis and also highlight the phosphopantetheine tunnel for tethering of the substrate to the nonribosomal peptide synthetase module during assembly line biosynthesis of the siderophore.
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U2 - 10.1021/acs.biochem.6b00735
DO - 10.1021/acs.biochem.6b00735
M3 - Article
C2 - 27601130
AN - SCOPUS:84989234534
SN - 0006-2960
VL - 55
SP - 5423
EP - 5433
JO - Biochemistry
JF - Biochemistry
IS - 38
ER -