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Microsomal glutathione transferase 1 exhibits one-third-of-the-sites-reactivity towards glutathione

Johan Ålander, Johan Lengqvist, Peter J Holm, Richard Svensson, Pascal Gerbaux, Robert H.H. van den Heuvel, Hans Hebert, William Griffiths Orcid Logo, Richard N Armstrong, Ralf Morgenstern

Archives of Biochemistry and Biophysics, Volume: 487, Issue: 1, Pages: 42 - 48

Swansea University Author: William Griffiths Orcid Logo

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DOI (Published version): 10.1016/j.abb.2009.04.009

Abstract

The trimeric membrane protein microsomal glutathione transferase 1 (MGST1) possesses glutathione transferase and peroxidase activity. Previous data indicated one active site/trimer whereas structural data suggests three GSH-binding sites. Here we have determined ligand interactions of MGST1 by sever...

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Published in: Archives of Biochemistry and Biophysics
ISSN: 0003-9861
Published: 2009
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URI: https://cronfa.swan.ac.uk/Record/cronfa10949
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first_indexed 2013-07-23T12:04:41Z
last_indexed 2018-02-09T04:40:13Z
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spelling 2011-10-01T00:00:00.0000000 v2 10949 2012-06-05 Microsomal glutathione transferase 1 exhibits one-third-of-the-sites-reactivity towards glutathione 3316b1d1b524be1831790933eed1c26e 0000-0002-4129-6616 William Griffiths William Griffiths true false 2012-06-05 BMS The trimeric membrane protein microsomal glutathione transferase 1 (MGST1) possesses glutathione transferase and peroxidase activity. Previous data indicated one active site/trimer whereas structural data suggests three GSH-binding sites. Here we have determined ligand interactions of MGST1 by several techniques. Nanoelectrospray mass spectrometry of native MGST1 revealed binding of three GSH molecules/trimer and equilibrium dialysis showed three product molecules/trimer (Kd = 320 ± 50 μM). All three product molecules could be competed out with GSH. Reinvestigation of GSH-binding showed one high affinity site per trimer, consistent with earlier data. Using single turnover stopped flow kinetic measurements, Kd could be determined for a low affinity GSH-binding site (2.5 ± 0.5 mM). Thus we can reconcile previous observations and show here that MGST1 contains three active sites with different affinities for GSH and that only the high affinity site is catalytically competent. Journal Article Archives of Biochemistry and Biophysics 487 1 42 48 0003-9861 31 12 2009 2009-12-31 10.1016/j.abb.2009.04.009 COLLEGE NANME Biomedical Sciences COLLEGE CODE BMS Swansea University 2011-10-01T00:00:00.0000000 2012-06-05T16:39:57.4585246 Faculty of Medicine, Health and Life Sciences Swansea University Medical School - Medicine Johan Ålander 1 Johan Lengqvist 2 Peter J Holm 3 Richard Svensson 4 Pascal Gerbaux 5 Robert H.H. van den Heuvel 6 Hans Hebert 7 William Griffiths 0000-0002-4129-6616 8 Richard N Armstrong 9 Ralf Morgenstern 10
title Microsomal glutathione transferase 1 exhibits one-third-of-the-sites-reactivity towards glutathione
spellingShingle Microsomal glutathione transferase 1 exhibits one-third-of-the-sites-reactivity towards glutathione
William Griffiths
title_short Microsomal glutathione transferase 1 exhibits one-third-of-the-sites-reactivity towards glutathione
title_full Microsomal glutathione transferase 1 exhibits one-third-of-the-sites-reactivity towards glutathione
title_fullStr Microsomal glutathione transferase 1 exhibits one-third-of-the-sites-reactivity towards glutathione
title_full_unstemmed Microsomal glutathione transferase 1 exhibits one-third-of-the-sites-reactivity towards glutathione
title_sort Microsomal glutathione transferase 1 exhibits one-third-of-the-sites-reactivity towards glutathione
author_id_str_mv 3316b1d1b524be1831790933eed1c26e
author_id_fullname_str_mv 3316b1d1b524be1831790933eed1c26e_***_William Griffiths
author William Griffiths
author2 Johan Ålander
Johan Lengqvist
Peter J Holm
Richard Svensson
Pascal Gerbaux
Robert H.H. van den Heuvel
Hans Hebert
William Griffiths
Richard N Armstrong
Ralf Morgenstern
format Journal article
container_title Archives of Biochemistry and Biophysics
container_volume 487
container_issue 1
container_start_page 42
publishDate 2009
institution Swansea University
issn 0003-9861
doi_str_mv 10.1016/j.abb.2009.04.009
college_str Faculty of Medicine, Health and Life Sciences
hierarchytype
hierarchy_top_id facultyofmedicinehealthandlifesciences
hierarchy_top_title Faculty of Medicine, Health and Life Sciences
hierarchy_parent_id facultyofmedicinehealthandlifesciences
hierarchy_parent_title Faculty of Medicine, Health and Life Sciences
department_str Swansea University Medical School - Medicine{{{_:::_}}}Faculty of Medicine, Health and Life Sciences{{{_:::_}}}Swansea University Medical School - Medicine
document_store_str 0
active_str 0
description The trimeric membrane protein microsomal glutathione transferase 1 (MGST1) possesses glutathione transferase and peroxidase activity. Previous data indicated one active site/trimer whereas structural data suggests three GSH-binding sites. Here we have determined ligand interactions of MGST1 by several techniques. Nanoelectrospray mass spectrometry of native MGST1 revealed binding of three GSH molecules/trimer and equilibrium dialysis showed three product molecules/trimer (Kd = 320 ± 50 μM). All three product molecules could be competed out with GSH. Reinvestigation of GSH-binding showed one high affinity site per trimer, consistent with earlier data. Using single turnover stopped flow kinetic measurements, Kd could be determined for a low affinity GSH-binding site (2.5 ± 0.5 mM). Thus we can reconcile previous observations and show here that MGST1 contains three active sites with different affinities for GSH and that only the high affinity site is catalytically competent.
published_date 2009-12-31T03:12:30Z
_version_ 1763750078808326144
score 11.016235

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