Holo structure and steady state kinetics of the thiazolinyl imine reductases for siderophore biosynthesis

Kathleen M. Meneely; Trey A. Ronnebaum; Andrew P. Riley; Thomas E. Prisinzano; Audrey L. Lamb

doi:10.1021/acs.biochem.6b00735

Holo structure and steady state kinetics of the thiazolinyl imine reductases for siderophore biosynthesis

Kathleen M. Meneely
, Trey A. Ronnebaum
, Andrew P. Riley
, Thomas E. Prisinzano
, Audrey L. Lamb

Research output: Contribution to journal › Article › peer-review

16 Scopus citations

Abstract

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.

Original language	English
Pages (from-to)	5423-5433
Number of pages	11
Journal	Biochemistry
Volume	55
Issue number	38
DOIs	https://doi.org/10.1021/acs.biochem.6b00735
State	Published - Sep 27 2016

Bibliographical note

Publisher Copyright:
© 2016 American Chemical Society.

Funding

This publication was made possible by funds from National Institutes of Health (NIH) Grants R01 AI77725 and K02 AI093675 from the National Institute for Allergy and Infectious Disease (A.L.L.), National Science Foundation Grant (NSF) CHE-1403293 (A.L.L.), and Graduate Training Program in Dynamic Aspects of Chemical Biology NIH Grant T32 GM008545 (T.A.R. and A.P.R.). Support for NMR instrumentation was provided by NIH Shared Instrumentation Grant S01 RR024664 and NSF Major Research Instrumentation Grant 0320648. Diffraction data were collected at the Stanford Synchrotron Radiation Laboratory (SSRL), a national user facility operated by Stanford University on behalf of the U.S. Department of Energy, Office of Biological and Environmental Research, and by the National Institutes of Health, National Center for Research Resources, Biomedical Technology Program, and the National Institute of General Medical Sciences.

Funders	Funder number
Biological and Environmental Research
National Institutes of Health (NIH)
U.S. Department of Energy Oak Ridge National Laboratory U.S. Department of Energy National Science Foundation National Energy Research Scientific Computing Center
Biomedical Research Technology Program
National Institute of General Medical Sciences	T32GM008545
U.S. Department of Energy Chinese Academy of Sciences Guangzhou Municipal Science and Technology Project Oak Ridge National Laboratory Extreme Science and Engineering Discovery Environment National Science Foundation National Energy Research Scientific Computing Center National Natural Science Foundation of China	0320648, S01 RR024664, CHE-1403293, T32 GM008545, 1403293
Division of Microbiology and Infectious Diseases, National Institute of Allergy and Infectious Diseases	K02AI093675, R01AI077725
National Center for Research Resources	S10RR024664

ASJC Scopus subject areas

Biochemistry

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10.1021/acs.biochem.6b00735

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title = "Holo structure and steady state kinetics of the thiazolinyl imine reductases for siderophore biosynthesis",

abstract = "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 {\AA} resolution and a holo structure to 1.28 {\AA} 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.",

author = "Meneely, \{Kathleen M.\} and Ronnebaum, \{Trey A.\} and Riley, \{Andrew P.\} and Prisinzano, \{Thomas E.\} and Lamb, \{Audrey L.\}",

note = "Publisher Copyright: {\textcopyright} 2016 American Chemical Society.",

year = "2016",

month = sep,

day = "27",

doi = "10.1021/acs.biochem.6b00735",

language = "English",

volume = "55",

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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.

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.

UR - https://www.scopus.com/pages/publications/84989234534

UR - https://www.scopus.com/pages/publications/84989234534#tab=citedBy

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 -

Holo structure and steady state kinetics of the thiazolinyl imine reductases for siderophore biosynthesis

Abstract

Bibliographical note

Funding

ASJC Scopus subject areas

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