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Use of complementary cation and anion heavy-atom salt derivatives to solve the structure of cytochrome P450 46A1
Human cytochrome P450 46A1 (CYP46A1) is one of the key enzymes in cholesterol homeostasis in the brain. The crystallization and heavy-atom structure solution of an active truncated CYP46A1 in complex with the high-affinity substrate analogue cholesterol-3-sulfate (CH-3S) is reported. The 2.6 Å struc...
Autores principales: | , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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International Union of Crystallography
2008
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2467524/ https://www.ncbi.nlm.nih.gov/pubmed/18453684 http://dx.doi.org/10.1107/S0907444908004046 |
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author | White, Mark Andrew Mast, Natalia Bjorkhem, Ingemar Johnson, Eric F. Stout, C. David Pikuleva, Irina A. |
author_facet | White, Mark Andrew Mast, Natalia Bjorkhem, Ingemar Johnson, Eric F. Stout, C. David Pikuleva, Irina A. |
author_sort | White, Mark Andrew |
collection | PubMed |
description | Human cytochrome P450 46A1 (CYP46A1) is one of the key enzymes in cholesterol homeostasis in the brain. The crystallization and heavy-atom structure solution of an active truncated CYP46A1 in complex with the high-affinity substrate analogue cholesterol-3-sulfate (CH-3S) is reported. The 2.6 Å structure of CYP46A1–CH-3S was solved using both anion and cation heavy-atom salts. In addition to the native anomalous signal from the haem iron, an NaI anion halide salt derivative and a complementary CsCl alkali-metal cation salt derivative were used. The general implications of the use of halide and alkali-metal quick soaks are discussed. The importance of using isoionic strength buffers, the titration of heavy-atom salts into different ionic species and the role of concentration are considered. It was observed that cation/anion-binding sites will occasionally overlap, which could negatively impact upon mixed RbBr soaks used for multiple anomalous scatterer MAD (MMAD). The use of complementary cation and anion heavy-atom salt derivatives is a convenient and powerful tool for MIR(AS) structure solution. |
format | Text |
id | pubmed-2467524 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2008 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
spelling | pubmed-24675242009-03-05 Use of complementary cation and anion heavy-atom salt derivatives to solve the structure of cytochrome P450 46A1 White, Mark Andrew Mast, Natalia Bjorkhem, Ingemar Johnson, Eric F. Stout, C. David Pikuleva, Irina A. Acta Crystallogr D Biol Crystallogr Research Papers Human cytochrome P450 46A1 (CYP46A1) is one of the key enzymes in cholesterol homeostasis in the brain. The crystallization and heavy-atom structure solution of an active truncated CYP46A1 in complex with the high-affinity substrate analogue cholesterol-3-sulfate (CH-3S) is reported. The 2.6 Å structure of CYP46A1–CH-3S was solved using both anion and cation heavy-atom salts. In addition to the native anomalous signal from the haem iron, an NaI anion halide salt derivative and a complementary CsCl alkali-metal cation salt derivative were used. The general implications of the use of halide and alkali-metal quick soaks are discussed. The importance of using isoionic strength buffers, the titration of heavy-atom salts into different ionic species and the role of concentration are considered. It was observed that cation/anion-binding sites will occasionally overlap, which could negatively impact upon mixed RbBr soaks used for multiple anomalous scatterer MAD (MMAD). The use of complementary cation and anion heavy-atom salt derivatives is a convenient and powerful tool for MIR(AS) structure solution. International Union of Crystallography 2008-05-01 2008-04-19 /pmc/articles/PMC2467524/ /pubmed/18453684 http://dx.doi.org/10.1107/S0907444908004046 Text en © International Union of Crystallography 2008 http://journals.iucr.org/services/termsofuse.html This is an open-access article distributed under the terms described at http://journals.iucr.org/services/termsofuse.html. |
spellingShingle | Research Papers White, Mark Andrew Mast, Natalia Bjorkhem, Ingemar Johnson, Eric F. Stout, C. David Pikuleva, Irina A. Use of complementary cation and anion heavy-atom salt derivatives to solve the structure of cytochrome P450 46A1 |
title | Use of complementary cation and anion heavy-atom salt derivatives to solve the structure of cytochrome P450 46A1 |
title_full | Use of complementary cation and anion heavy-atom salt derivatives to solve the structure of cytochrome P450 46A1 |
title_fullStr | Use of complementary cation and anion heavy-atom salt derivatives to solve the structure of cytochrome P450 46A1 |
title_full_unstemmed | Use of complementary cation and anion heavy-atom salt derivatives to solve the structure of cytochrome P450 46A1 |
title_short | Use of complementary cation and anion heavy-atom salt derivatives to solve the structure of cytochrome P450 46A1 |
title_sort | use of complementary cation and anion heavy-atom salt derivatives to solve the structure of cytochrome p450 46a1 |
topic | Research Papers |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2467524/ https://www.ncbi.nlm.nih.gov/pubmed/18453684 http://dx.doi.org/10.1107/S0907444908004046 |
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