Experimental correlation of substrate position with reaction outcome in the aliphatic halogenase, SyrB2

Ryan J. Martinie, Jovan Livada, Wei Chen Chang, Michael T. Green, Carsten Krebs, Joseph M. Bollinger, Jr., Alexey Silakov

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

The iron(II)- and 2-(oxo)glutarate-dependent (Fe/2OG) oxygenases catalyze an array of challenging transformations, but how individual members of the enzyme family direct different outcomes is poorly understood. The Fe/2OG halogenase, SyrB2, chlorinates C4 of its native substrate, l-threonine appended to the carrier protein, SyrB1, but hydroxylates C5 of l-norvaline and, to a lesser extent, C4 of l-aminobutyric acid when SyrB1 presents these non-native amino acids. To test the hypothesis that positioning of the targeted carbon dictates the outcome, we defined the positions of these three substrates by measuring hyperfine couplings between substrate deuterium atoms and the stable, EPR-active iron-nitrosyl adduct, a surrogate for reaction intermediates. The Fe- 2 H distances and N-Fe- 2 H angles, which vary from 4.2 Å and 85° for threonine to 3.4 Å and 65° for norvaline, rationalize the trends in reactivity. This experimental correlation of position to outcome should aid in judging from structural data on other Fe/2OG enzymes whether they suppress hydroxylation or form hydroxylated intermediates on the pathways to other outcomes.

Original languageEnglish (US)
Pages (from-to)6912-6919
Number of pages8
JournalJournal of the American Chemical Society
Volume137
Issue number21
DOIs
StatePublished - Jun 3 2015

Fingerprint

Threonine
Glutarates
Aminobutyrates
Oxygenases
Deuterium
Substrates
Enzymes
Hydroxylation
Iron
Reaction intermediates
Carrier Proteins
Carbon
Amino Acids
Paramagnetic resonance
Amino acids
Atoms
Acids
norvaline
dinitrosyl iron complex

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

@article{0a87687e56a74a899e7adc784d7c9e22,
title = "Experimental correlation of substrate position with reaction outcome in the aliphatic halogenase, SyrB2",
abstract = "The iron(II)- and 2-(oxo)glutarate-dependent (Fe/2OG) oxygenases catalyze an array of challenging transformations, but how individual members of the enzyme family direct different outcomes is poorly understood. The Fe/2OG halogenase, SyrB2, chlorinates C4 of its native substrate, l-threonine appended to the carrier protein, SyrB1, but hydroxylates C5 of l-norvaline and, to a lesser extent, C4 of l-aminobutyric acid when SyrB1 presents these non-native amino acids. To test the hypothesis that positioning of the targeted carbon dictates the outcome, we defined the positions of these three substrates by measuring hyperfine couplings between substrate deuterium atoms and the stable, EPR-active iron-nitrosyl adduct, a surrogate for reaction intermediates. The Fe- 2 H distances and N-Fe- 2 H angles, which vary from 4.2 {\AA} and 85° for threonine to 3.4 {\AA} and 65° for norvaline, rationalize the trends in reactivity. This experimental correlation of position to outcome should aid in judging from structural data on other Fe/2OG enzymes whether they suppress hydroxylation or form hydroxylated intermediates on the pathways to other outcomes.",
author = "Martinie, {Ryan J.} and Jovan Livada and Chang, {Wei Chen} and Green, {Michael T.} and Carsten Krebs and {Bollinger, Jr.}, {Joseph M.} and Alexey Silakov",
year = "2015",
month = "6",
day = "3",
doi = "10.1021/jacs.5b03370",
language = "English (US)",
volume = "137",
pages = "6912--6919",
journal = "Journal of the American Chemical Society",
issn = "0002-7863",
publisher = "American Chemical Society",
number = "21",

}

Experimental correlation of substrate position with reaction outcome in the aliphatic halogenase, SyrB2. / Martinie, Ryan J.; Livada, Jovan; Chang, Wei Chen; Green, Michael T.; Krebs, Carsten; Bollinger, Jr., Joseph M.; Silakov, Alexey.

In: Journal of the American Chemical Society, Vol. 137, No. 21, 03.06.2015, p. 6912-6919.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Experimental correlation of substrate position with reaction outcome in the aliphatic halogenase, SyrB2

AU - Martinie, Ryan J.

AU - Livada, Jovan

AU - Chang, Wei Chen

AU - Green, Michael T.

AU - Krebs, Carsten

AU - Bollinger, Jr., Joseph M.

AU - Silakov, Alexey

PY - 2015/6/3

Y1 - 2015/6/3

N2 - The iron(II)- and 2-(oxo)glutarate-dependent (Fe/2OG) oxygenases catalyze an array of challenging transformations, but how individual members of the enzyme family direct different outcomes is poorly understood. The Fe/2OG halogenase, SyrB2, chlorinates C4 of its native substrate, l-threonine appended to the carrier protein, SyrB1, but hydroxylates C5 of l-norvaline and, to a lesser extent, C4 of l-aminobutyric acid when SyrB1 presents these non-native amino acids. To test the hypothesis that positioning of the targeted carbon dictates the outcome, we defined the positions of these three substrates by measuring hyperfine couplings between substrate deuterium atoms and the stable, EPR-active iron-nitrosyl adduct, a surrogate for reaction intermediates. The Fe- 2 H distances and N-Fe- 2 H angles, which vary from 4.2 Å and 85° for threonine to 3.4 Å and 65° for norvaline, rationalize the trends in reactivity. This experimental correlation of position to outcome should aid in judging from structural data on other Fe/2OG enzymes whether they suppress hydroxylation or form hydroxylated intermediates on the pathways to other outcomes.

AB - The iron(II)- and 2-(oxo)glutarate-dependent (Fe/2OG) oxygenases catalyze an array of challenging transformations, but how individual members of the enzyme family direct different outcomes is poorly understood. The Fe/2OG halogenase, SyrB2, chlorinates C4 of its native substrate, l-threonine appended to the carrier protein, SyrB1, but hydroxylates C5 of l-norvaline and, to a lesser extent, C4 of l-aminobutyric acid when SyrB1 presents these non-native amino acids. To test the hypothesis that positioning of the targeted carbon dictates the outcome, we defined the positions of these three substrates by measuring hyperfine couplings between substrate deuterium atoms and the stable, EPR-active iron-nitrosyl adduct, a surrogate for reaction intermediates. The Fe- 2 H distances and N-Fe- 2 H angles, which vary from 4.2 Å and 85° for threonine to 3.4 Å and 65° for norvaline, rationalize the trends in reactivity. This experimental correlation of position to outcome should aid in judging from structural data on other Fe/2OG enzymes whether they suppress hydroxylation or form hydroxylated intermediates on the pathways to other outcomes.

UR - http://www.scopus.com/inward/record.url?scp=84930670202&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84930670202&partnerID=8YFLogxK

U2 - 10.1021/jacs.5b03370

DO - 10.1021/jacs.5b03370

M3 - Article

C2 - 25965587

AN - SCOPUS:84930670202

VL - 137

SP - 6912

EP - 6919

JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

SN - 0002-7863

IS - 21

ER -