Crystallization and preliminary x-ray diffraction analysis of P450terp and the hemoprotein domain of P450BM-3, enzymes belonging to two distinct classes of the cytochrome P450 superfamily

Sekhar S. Boddupalli, Charles A. Hasemann, K. G. Ravichandran, Jui Yun Lu, Elizabeth J. Goldsmith, Johann Deisenhofer, Julian A. Peterson

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Abstract

Cytochromes P450 are members of a superfamily of hemoproteins that are involved in the metabolism of various physiologic and xenobiotic organic compounds. This superfamily of proteins can be divided into two classes based on the electron donor proximal to the P450: an iron-sulfur protein for class I P450s or a flavoprotein for class II. The only known tertiary structure of any of the cytochromes P450 is that of P450cam, a class I soluble enzyme isolated from Pseudomonas putida (product of the CYP101 gene). To understand the details of the structure-function relationships within and between the two classes, structural studies on additional cytochromes P450 are crucial. We report here characterization of the crystal forms of two soluble, bacterial enzymes: cytochrome P450trep [class I enzyme from a Pseudomonas species (product of CYP108 gene)] and the hemoprotein domain of cytochrome P450BM-3 [class II enzyme from Bacillus megaterium (product of the CYP102 gene)]. The crystals of cytochrome P450terp are hexagonal and belong to the space group P6122 (or its enantiomorph, P6522) with unit cell dimensions a = b = 68.9 Å and c = 458.7 Å. The crystals of the hemoprotein domain of cytochrome P450BM-3 are monoclinic and belong to the space group P21 with unit cell dimensions a = 59.4 Å, b = 154.0 Å, c = 62.2 Å, and β = 94.7°. Diffraction data for the crystals of these two proteins were obtained to a resolution better than 2.2 Å. Assuming the presence of two molecules in the asymmetric unit for the hemoprotein domain of P450BM-3 and one molecule for P450terp, the calculated values of Vm are 2.6 and 3.3 Å3/Da, respectively.

Original languageEnglish (US)
Pages (from-to)5567-5571
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume89
Issue number12
StatePublished - 1992

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Cytochromes
Crystallization
Cytochrome P-450 Enzyme System
Camphor 5-Monooxygenase
X-Rays
Enzymes
Iron-Sulfur Proteins
Genes
Bacillus megaterium
Flavoproteins
Pseudomonas putida
Xenobiotics
Pseudomonas
Proteins
Electrons
terpineol hydroxylase

Keywords

  • Bacterial P450
  • P450 structure

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

@article{838b88c8ca6f40589ffdb23d16907f29,
title = "Crystallization and preliminary x-ray diffraction analysis of P450terp and the hemoprotein domain of P450BM-3, enzymes belonging to two distinct classes of the cytochrome P450 superfamily",
abstract = "Cytochromes P450 are members of a superfamily of hemoproteins that are involved in the metabolism of various physiologic and xenobiotic organic compounds. This superfamily of proteins can be divided into two classes based on the electron donor proximal to the P450: an iron-sulfur protein for class I P450s or a flavoprotein for class II. The only known tertiary structure of any of the cytochromes P450 is that of P450cam, a class I soluble enzyme isolated from Pseudomonas putida (product of the CYP101 gene). To understand the details of the structure-function relationships within and between the two classes, structural studies on additional cytochromes P450 are crucial. We report here characterization of the crystal forms of two soluble, bacterial enzymes: cytochrome P450trep [class I enzyme from a Pseudomonas species (product of CYP108 gene)] and the hemoprotein domain of cytochrome P450BM-3 [class II enzyme from Bacillus megaterium (product of the CYP102 gene)]. The crystals of cytochrome P450terp are hexagonal and belong to the space group P6122 (or its enantiomorph, P6522) with unit cell dimensions a = b = 68.9 {\AA} and c = 458.7 {\AA}. The crystals of the hemoprotein domain of cytochrome P450BM-3 are monoclinic and belong to the space group P21 with unit cell dimensions a = 59.4 {\AA}, b = 154.0 {\AA}, c = 62.2 {\AA}, and β = 94.7°. Diffraction data for the crystals of these two proteins were obtained to a resolution better than 2.2 {\AA}. Assuming the presence of two molecules in the asymmetric unit for the hemoprotein domain of P450BM-3 and one molecule for P450terp, the calculated values of Vm are 2.6 and 3.3 {\AA}3/Da, respectively.",
keywords = "Bacterial P450, P450 structure",
author = "Boddupalli, {Sekhar S.} and Hasemann, {Charles A.} and Ravichandran, {K. G.} and Lu, {Jui Yun} and Goldsmith, {Elizabeth J.} and Johann Deisenhofer and Peterson, {Julian A.}",
year = "1992",
language = "English (US)",
volume = "89",
pages = "5567--5571",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "12",

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TY - JOUR

T1 - Crystallization and preliminary x-ray diffraction analysis of P450terp and the hemoprotein domain of P450BM-3, enzymes belonging to two distinct classes of the cytochrome P450 superfamily

AU - Boddupalli, Sekhar S.

AU - Hasemann, Charles A.

AU - Ravichandran, K. G.

AU - Lu, Jui Yun

AU - Goldsmith, Elizabeth J.

AU - Deisenhofer, Johann

AU - Peterson, Julian A.

PY - 1992

Y1 - 1992

N2 - Cytochromes P450 are members of a superfamily of hemoproteins that are involved in the metabolism of various physiologic and xenobiotic organic compounds. This superfamily of proteins can be divided into two classes based on the electron donor proximal to the P450: an iron-sulfur protein for class I P450s or a flavoprotein for class II. The only known tertiary structure of any of the cytochromes P450 is that of P450cam, a class I soluble enzyme isolated from Pseudomonas putida (product of the CYP101 gene). To understand the details of the structure-function relationships within and between the two classes, structural studies on additional cytochromes P450 are crucial. We report here characterization of the crystal forms of two soluble, bacterial enzymes: cytochrome P450trep [class I enzyme from a Pseudomonas species (product of CYP108 gene)] and the hemoprotein domain of cytochrome P450BM-3 [class II enzyme from Bacillus megaterium (product of the CYP102 gene)]. The crystals of cytochrome P450terp are hexagonal and belong to the space group P6122 (or its enantiomorph, P6522) with unit cell dimensions a = b = 68.9 Å and c = 458.7 Å. The crystals of the hemoprotein domain of cytochrome P450BM-3 are monoclinic and belong to the space group P21 with unit cell dimensions a = 59.4 Å, b = 154.0 Å, c = 62.2 Å, and β = 94.7°. Diffraction data for the crystals of these two proteins were obtained to a resolution better than 2.2 Å. Assuming the presence of two molecules in the asymmetric unit for the hemoprotein domain of P450BM-3 and one molecule for P450terp, the calculated values of Vm are 2.6 and 3.3 Å3/Da, respectively.

AB - Cytochromes P450 are members of a superfamily of hemoproteins that are involved in the metabolism of various physiologic and xenobiotic organic compounds. This superfamily of proteins can be divided into two classes based on the electron donor proximal to the P450: an iron-sulfur protein for class I P450s or a flavoprotein for class II. The only known tertiary structure of any of the cytochromes P450 is that of P450cam, a class I soluble enzyme isolated from Pseudomonas putida (product of the CYP101 gene). To understand the details of the structure-function relationships within and between the two classes, structural studies on additional cytochromes P450 are crucial. We report here characterization of the crystal forms of two soluble, bacterial enzymes: cytochrome P450trep [class I enzyme from a Pseudomonas species (product of CYP108 gene)] and the hemoprotein domain of cytochrome P450BM-3 [class II enzyme from Bacillus megaterium (product of the CYP102 gene)]. The crystals of cytochrome P450terp are hexagonal and belong to the space group P6122 (or its enantiomorph, P6522) with unit cell dimensions a = b = 68.9 Å and c = 458.7 Å. The crystals of the hemoprotein domain of cytochrome P450BM-3 are monoclinic and belong to the space group P21 with unit cell dimensions a = 59.4 Å, b = 154.0 Å, c = 62.2 Å, and β = 94.7°. Diffraction data for the crystals of these two proteins were obtained to a resolution better than 2.2 Å. Assuming the presence of two molecules in the asymmetric unit for the hemoprotein domain of P450BM-3 and one molecule for P450terp, the calculated values of Vm are 2.6 and 3.3 Å3/Da, respectively.

KW - Bacterial P450

KW - P450 structure

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M3 - Article

VL - 89

SP - 5567

EP - 5571

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 12

ER -