Cargando…
X-Ray Structure and Mutagenesis Studies of the N-Isopropylammelide Isopropylaminohydrolase, AtzC
The N-isopropylammelide isopropylaminohydrolase from Pseudomonas sp. strain ADP, AtzC, provides the third hydrolytic step in the mineralization of s-triazine herbicides, such as atrazine. We obtained the X-ray crystal structure of AtzC at 1.84 Å with a weak inhibitor bound in the active site and the...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2015
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4577212/ https://www.ncbi.nlm.nih.gov/pubmed/26390431 http://dx.doi.org/10.1371/journal.pone.0137700 |
_version_ | 1782390952260796416 |
---|---|
author | Balotra, Sahil Warden, Andrew C. Newman, Janet Briggs, Lyndall J. Scott, Colin Peat, Thomas S. |
author_facet | Balotra, Sahil Warden, Andrew C. Newman, Janet Briggs, Lyndall J. Scott, Colin Peat, Thomas S. |
author_sort | Balotra, Sahil |
collection | PubMed |
description | The N-isopropylammelide isopropylaminohydrolase from Pseudomonas sp. strain ADP, AtzC, provides the third hydrolytic step in the mineralization of s-triazine herbicides, such as atrazine. We obtained the X-ray crystal structure of AtzC at 1.84 Å with a weak inhibitor bound in the active site and then used a combination of in silico docking and site-directed mutagenesis to understand the interactions between AtzC and its substrate, isopropylammelide. The substitution of an active site histidine residue (His249) for an alanine abolished the enzyme’s catalytic activity. We propose a plausible catalytic mechanism, consistent with the biochemical and crystallographic data obtained that is similar to that found in carbonic anhydrase and other members of subtype III of the amidohydrolase family |
format | Online Article Text |
id | pubmed-4577212 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-45772122015-09-25 X-Ray Structure and Mutagenesis Studies of the N-Isopropylammelide Isopropylaminohydrolase, AtzC Balotra, Sahil Warden, Andrew C. Newman, Janet Briggs, Lyndall J. Scott, Colin Peat, Thomas S. PLoS One Research Article The N-isopropylammelide isopropylaminohydrolase from Pseudomonas sp. strain ADP, AtzC, provides the third hydrolytic step in the mineralization of s-triazine herbicides, such as atrazine. We obtained the X-ray crystal structure of AtzC at 1.84 Å with a weak inhibitor bound in the active site and then used a combination of in silico docking and site-directed mutagenesis to understand the interactions between AtzC and its substrate, isopropylammelide. The substitution of an active site histidine residue (His249) for an alanine abolished the enzyme’s catalytic activity. We propose a plausible catalytic mechanism, consistent with the biochemical and crystallographic data obtained that is similar to that found in carbonic anhydrase and other members of subtype III of the amidohydrolase family Public Library of Science 2015-09-21 /pmc/articles/PMC4577212/ /pubmed/26390431 http://dx.doi.org/10.1371/journal.pone.0137700 Text en © 2015 Balotra et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Balotra, Sahil Warden, Andrew C. Newman, Janet Briggs, Lyndall J. Scott, Colin Peat, Thomas S. X-Ray Structure and Mutagenesis Studies of the N-Isopropylammelide Isopropylaminohydrolase, AtzC |
title | X-Ray Structure and Mutagenesis Studies of the N-Isopropylammelide Isopropylaminohydrolase, AtzC |
title_full | X-Ray Structure and Mutagenesis Studies of the N-Isopropylammelide Isopropylaminohydrolase, AtzC |
title_fullStr | X-Ray Structure and Mutagenesis Studies of the N-Isopropylammelide Isopropylaminohydrolase, AtzC |
title_full_unstemmed | X-Ray Structure and Mutagenesis Studies of the N-Isopropylammelide Isopropylaminohydrolase, AtzC |
title_short | X-Ray Structure and Mutagenesis Studies of the N-Isopropylammelide Isopropylaminohydrolase, AtzC |
title_sort | x-ray structure and mutagenesis studies of the n-isopropylammelide isopropylaminohydrolase, atzc |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4577212/ https://www.ncbi.nlm.nih.gov/pubmed/26390431 http://dx.doi.org/10.1371/journal.pone.0137700 |
work_keys_str_mv | AT balotrasahil xraystructureandmutagenesisstudiesofthenisopropylammelideisopropylaminohydrolaseatzc AT wardenandrewc xraystructureandmutagenesisstudiesofthenisopropylammelideisopropylaminohydrolaseatzc AT newmanjanet xraystructureandmutagenesisstudiesofthenisopropylammelideisopropylaminohydrolaseatzc AT briggslyndallj xraystructureandmutagenesisstudiesofthenisopropylammelideisopropylaminohydrolaseatzc AT scottcolin xraystructureandmutagenesisstudiesofthenisopropylammelideisopropylaminohydrolaseatzc AT peatthomass xraystructureandmutagenesisstudiesofthenisopropylammelideisopropylaminohydrolaseatzc |