Abstract
Affinity labeling, using reactive analogs of natural ligands, provides a powerful approach to probe for the existence of cooperative interactions between subunits as well as to locate specific xenobiotic binding sites in glutathione 5-transferases (GST). Thus, 3β-(iodoacetoxy)-dehydroisoandrosterone (3β-IDA) and 17-(iodoacetoxy)estradiol-3-sulfate (17β-IES) function as affinity labels of the nonsubstrate steroid binding site of rat liver GST, isozyme 1-1. Modification of Cys17 or Cys111 on Subunit A prevents reaction of the same cysteine on Subunit B. Interactions between the subunits of both the 4-4 and 1-1 isozymes are also illustrated by photoaffinity labeling of the active sites of these homodimers by glutathionyl S[4-(succinimidyl)-benzophenone], an analog of the product of glutathione and xenobiotic substrate.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 159-162 |
| Number of pages | 4 |
| Journal | Chemico-Biological Interactions |
| Volume | 133 |
| Issue number | 1-3 |
| State | Published - Feb 28 2001 |
| Externally published | Yes |
Keywords
- Affinity labeling
- Glutathione S-transferase
- Subunit interaction
ASJC Scopus subject areas
- Toxicology
Other files and links
Fingerprint
Dive into the research topics of 'Probing subunit interactions in glutathione S-transferases using affinity labeling'. Together they form a unique fingerprint.Cite this
- APA
- Standard
- Harvard
- Vancouver
- Author
- BIBTEX
- RIS
In: Chemico-Biological Interactions, Vol. 133, No. 1-3, 28.02.2001, p. 159-162.
Research output: Contribution to journal › Article › peer-review
}
TY - JOUR
T1 - Probing subunit interactions in glutathione S-transferases using affinity labeling
AU - Colman, Roberta F.
AU - Barycki, Joseph J.
AU - Vargo, Melissa A.
AU - Wang, Jibo
N1 - Funding Information: This work was supported by a grant from the US Army Medical Research and Material Command (Breast Cancer Research Program). Funding Information: The author would like to acknowledge the invaluable contributions by the postgraduate students and staff of the Protein Structure-Function Research Programme and Richard Armstrong (Vanderbilt University) to this work. Financial support was provided by Wits University, National Research Foundation, Alexander von Humboldt Foundation and FIRCA(NIH) grant TW00779. Funding Information: A.-M. Abdalla was supported by a scholarship from the Egyptian Government. This work was funded by grants from the Swedish Natural Science Research Council and the Carl Trygger Foundation for Scientific Research. Funding Information: This work was supported in part by SNF grant 3100-050602.97 to S.V. Funding Information: The Finnish studies were supported by the Academy of Finland, the Finnish Konkordia Foundation, and EVO funds from Kuopio University Hospital. The French study was supported by the Swiss Cancer League, the League against Cancer of Fribourg, the Cancer Research (Switzerland) and the Gustave–Roussy Institute (France). Funding Information: This work was supported by United States Public Health Service Grants CA-66561 and T32-GM-08550. Funding Information: This work was supported by the University of the Witwatersrand, the South African National Research Foundation and the Fogarty International Research Collaboration Award TW00779. Funding Information: This study is part of special projects supported by Italian Ministry of University and of Scientific and Technological Research, Italian National Research Council and Italian Ministry of Health. Funding Information: This work is supported by the DFG (grant Li 1-3, 1-4). Funding Information: The work presented here was partially supported by the Deutsche Forschungsge-meinschaft (SFB 475). Funding Information: The author would like to thank Ralph Hermanns for technical assistance and Professor Peter van Bladeren for the rat GST isoenzymes. This research was funded by the Dutch Cancer Society (project RUL017-1407). Funding Information: The ATPase activity of recombinant RalBP1 was only marginally stimulated by DNP-SG (by approximately 30%). This may indicate that the protein, as isolated from the bacterial lysate, is saturated with a tightly bound ligand and already near-maximally stimulated. Antibiotics used for plasmid selection are possible candidates for such ligands. This interpretation was supported by the fact that omitting chloramphenicol from the bacterial growth medium improved stimulation. Funding Information: This work was supported by grants from the U.S. National Cancer Institute (76420), the Howard Hughes Medical Institute, and the Oakland University Research Excellence Fund. Funding Information: I gratefully acknowledge Jim Parsons at the Vanderbilt University School of Medicine for the generation of the recombinant mutant plasmids. This work was supported by the University of the Witwatersrand, the South African National Research Foundation, the Fogarty International Collaboration Award TW00779, Grant GM30910 from the National Institutes of Health and the Alexander von Humboldt Foundation. Funding Information: The Andrew Mellon Foundation, the University of the Witwatersrand and the South African National Research Foundation for financial support. Funding Information: The author wishes to acknowledge the following collaborators, Richard N. Armstrong, Bryan Bernat and Richard Svensson in the kinetic studies and Hans Hebert in structural aspects. The studies were supported by the Swedish Cancer Society, The National Board for Laboratory Animals, Carl Tryggers Foundation and funds from Karolinska Insitutet. Funding Information: This work is supported by National Institute of Health Grants AI22531, AI31599, AR36308, ES06105, HL03208. Funding Information: Supported by the British Lung Foundation, MURST, Ministero del Lavoro e della Previdenza Sociale, Danish Research Academy and North Staffordshire Medical Institute. Funding Information: This work was supported by MURST-40%; grants telethon-E872, AIRC, EU (QRLT-1999-00739); Ministero Sanità. Funding Information: Support from the Swedish Medical Research Council (projects no 31X-12573), the Swedish Society of Medicine, the Swedish Cancer Society, Harald Jeansson’s and Harald and Greta Jeansson’s, and the Karolinska Institutet foundations are gratefully acknowledged. Funding Information: Financial support is acknowledged with grateful thanks from the International Programs in Chemical Sciences (IPICS), Uppsala University and the Research Board, University of Zimbabwe. Funding Information: We thank the staff of ESRF synchrotron, Grenoble and the SRS synchrotron, Daresbury for help with data collection. Financial support from ESRF for our visit to Grenoble is gratefully acknowledged. We also acknowledge Aventis Crop Science UK Ltd. for funding this project. Funding Information: Supported by The North Staffordshire Medical Institute and the McCall Foundation. Funding Information: This work was supported in part by NIH:NIEHS grant ES07804 (to PZ) and by a pilot grant from the University of Arkansas for Medical Science. Funding Information: Funding for this project was provided by grants from the National Institutes of Health (R01-EOS9427) and the U.S. Environmental Protection Agency (R 827441)– Funding Information: This work was supported in part by an Australian Research Council Project Grant and an Australian Research Council Senior Research Fellowship (to M.W. Parker), a National Research Council of Italy Grant (Target Project on Biotechnology, to G. Ricci), a National Health and Medical Research Council Postgraduate Research Scholarship and an International Centre for Diffraction Data Crystallography Scholarship (to A.J. Oakley) and an Australian Research Council Postdoctoral Fellowship (to J. Rossjohn). Funding Information: The research in our laboratory is supported by grants from The Swedish Medical Research Council and from AstraZeneca. Funding Information: This work was funded by Aventis Crop Science UK Ltd. Funding Information: This work was funded by a grant from the Association of International Cancer Research (99-041, awarded to JDH and MY). Simon A. Chanas thanks the MRC and Zeneca for a collaborative Ph.D. studentship. Funding Information: H.J.L. was supported by NIH grants 1R01CA66782 and 1R01CA73403. Funding Information: This work was supported by the Research Science Funds of Serbia, Grant c03E18. Funding Information: This work was supported in part by a grant from the Ministero dell’ Università e della Ricerca Scientifica e Tecnologica. Funding Information: We thank the Cancer Research Campaign, British Medical Association and British Lung Foundation for support. Funding Information: In part supported by grants GM32304 (YCA), CA77495 (SA), VA Merit Review (PZ), and CA55589 (SVS). Funding Information: The authors would like to acknowledge invaluable contributions of L.T. Laugh-lin, B. Bernat and Professor John Helman (Cornell University) to various aspects of this work. NIH Grants A142756 and GM30910 and the Training Program in Molecular Biophysics at Vanderbilt provided support for this work. Funding Information: We thank Pedro Miguel Santos for sending pPR9TT plasmid and E. coli strains used in this work and P. Di Nardo for checking the English. This work was supported in part by European Community contract no. QLK3-CT1999–00041. Funding Information: This work was supported in part by grant from the Ministero dell’ Università e della Ricerca Scientifica e Tecnologica. Funding Information: In part supported by grants CA27967 (YCA), CA77495 (SA), and ES07804 (PZ). Funding Information: This investigation received financial support from the UNDP:WORLD BANK: WHO Special Programme for Research and Training in Tropical Medicine (TDR0096 and TDR 980281). Funding Information: This work was supported by a grant from the National Science Foundation (MCB-9723308). Funding Information: This research was supported in part by National Institute of Environmental Health Sciences grant ES03127 to M.W.A.
PY - 2001/2/28
Y1 - 2001/2/28
N2 - Affinity labeling, using reactive analogs of natural ligands, provides a powerful approach to probe for the existence of cooperative interactions between subunits as well as to locate specific xenobiotic binding sites in glutathione 5-transferases (GST). Thus, 3β-(iodoacetoxy)-dehydroisoandrosterone (3β-IDA) and 17-(iodoacetoxy)estradiol-3-sulfate (17β-IES) function as affinity labels of the nonsubstrate steroid binding site of rat liver GST, isozyme 1-1. Modification of Cys17 or Cys111 on Subunit A prevents reaction of the same cysteine on Subunit B. Interactions between the subunits of both the 4-4 and 1-1 isozymes are also illustrated by photoaffinity labeling of the active sites of these homodimers by glutathionyl S[4-(succinimidyl)-benzophenone], an analog of the product of glutathione and xenobiotic substrate.
AB - Affinity labeling, using reactive analogs of natural ligands, provides a powerful approach to probe for the existence of cooperative interactions between subunits as well as to locate specific xenobiotic binding sites in glutathione 5-transferases (GST). Thus, 3β-(iodoacetoxy)-dehydroisoandrosterone (3β-IDA) and 17-(iodoacetoxy)estradiol-3-sulfate (17β-IES) function as affinity labels of the nonsubstrate steroid binding site of rat liver GST, isozyme 1-1. Modification of Cys17 or Cys111 on Subunit A prevents reaction of the same cysteine on Subunit B. Interactions between the subunits of both the 4-4 and 1-1 isozymes are also illustrated by photoaffinity labeling of the active sites of these homodimers by glutathionyl S[4-(succinimidyl)-benzophenone], an analog of the product of glutathione and xenobiotic substrate.
KW - Affinity labeling
KW - Glutathione S-transferase
KW - Subunit interaction
UR - http://www.scopus.com/inward/record.url?scp=0000336613&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0000336613&partnerID=8YFLogxK
M3 - Article
AN - SCOPUS:0000336613
SN - 0009-2797
VL - 133
SP - 159
EP - 162
JO - Chemico-Biological Interactions
JF - Chemico-Biological Interactions
IS - 1-3
ER -