Unveiling the crucial intermediates in androgen production

Piotr J. Mak, Michael C. Gregory, Ilia G. Denisov, Stephen G. Sligar, James R. Kincaid

Research output: Contribution to journalArticle

Abstract

Ablation of androgen production through surgery is one strategy against prostate cancer, with the current focus placed on pharmaceutical intervention to restrict androgen synthesis selectively, an endeavor that could benefit from the enhanced understanding of enzymatic mechanisms that derives from characterization of key reaction intermediates. The multifunctional cytochrome P450 17A1 (CYP17A1) first catalyzes the typical hydroxylation of its primary substrate, pregnenolone (PREG) and then also orchestrates a remarkable C17-C20 bond cleavage (lyase) reaction, converting the 17-hydroxypregnenolone initial product to dehydroepiandrosterone, a process representing the first committed step in the biosynthesis of androgens. Now, we report the capture and structural characterization of intermediates produced during this lyase step: an initial peroxo-anion intermediate, poised for nucleophilic attack on the C20 position by a substrate-associated H-bond, and the crucial ferric peroxo-hemiacetal intermediate that precedes carbon-carbon (C-C) bond cleavage. These studies provide a rare glimpse at the actual structural determinants of a chemical transformation that carries profound physiological consequences.

Original languageEnglish (US)
Pages (from-to)15856-15861
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume112
Issue number52
DOIs
StatePublished - Dec 29 2015

Fingerprint

Androgens
Lyases
Carbon
17-alpha-Hydroxypregnenolone
Pregnenolone
Dehydroepiandrosterone
Hydroxylation
Cytochrome P-450 Enzyme System
Anions
Prostatic Neoplasms
Pharmaceutical Preparations

Keywords

  • Cytochrome P450
  • Nanodiscs
  • Peroxo-hemiacetal
  • Resonance Raman spectroscopy
  • Steroids

ASJC Scopus subject areas

  • General

Cite this

Unveiling the crucial intermediates in androgen production. / Mak, Piotr J.; Gregory, Michael C.; Denisov, Ilia G.; Sligar, Stephen G.; Kincaid, James R.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 112, No. 52, 29.12.2015, p. 15856-15861.

Research output: Contribution to journalArticle

Mak, Piotr J. ; Gregory, Michael C. ; Denisov, Ilia G. ; Sligar, Stephen G. ; Kincaid, James R. / Unveiling the crucial intermediates in androgen production. In: Proceedings of the National Academy of Sciences of the United States of America. 2015 ; Vol. 112, No. 52. pp. 15856-15861.
@article{670c3e3b809e48ab907ba1b14222a7a9,
title = "Unveiling the crucial intermediates in androgen production",
abstract = "Ablation of androgen production through surgery is one strategy against prostate cancer, with the current focus placed on pharmaceutical intervention to restrict androgen synthesis selectively, an endeavor that could benefit from the enhanced understanding of enzymatic mechanisms that derives from characterization of key reaction intermediates. The multifunctional cytochrome P450 17A1 (CYP17A1) first catalyzes the typical hydroxylation of its primary substrate, pregnenolone (PREG) and then also orchestrates a remarkable C17-C20 bond cleavage (lyase) reaction, converting the 17-hydroxypregnenolone initial product to dehydroepiandrosterone, a process representing the first committed step in the biosynthesis of androgens. Now, we report the capture and structural characterization of intermediates produced during this lyase step: an initial peroxo-anion intermediate, poised for nucleophilic attack on the C20 position by a substrate-associated H-bond, and the crucial ferric peroxo-hemiacetal intermediate that precedes carbon-carbon (C-C) bond cleavage. These studies provide a rare glimpse at the actual structural determinants of a chemical transformation that carries profound physiological consequences.",
keywords = "Cytochrome P450, Nanodiscs, Peroxo-hemiacetal, Resonance Raman spectroscopy, Steroids",
author = "Mak, {Piotr J.} and Gregory, {Michael C.} and Denisov, {Ilia G.} and Sligar, {Stephen G.} and Kincaid, {James R.}",
year = "2015",
month = "12",
day = "29",
doi = "10.1073/pnas.1519376113",
language = "English (US)",
volume = "112",
pages = "15856--15861",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "52",

}

TY - JOUR

T1 - Unveiling the crucial intermediates in androgen production

AU - Mak, Piotr J.

AU - Gregory, Michael C.

AU - Denisov, Ilia G.

AU - Sligar, Stephen G.

AU - Kincaid, James R.

PY - 2015/12/29

Y1 - 2015/12/29

N2 - Ablation of androgen production through surgery is one strategy against prostate cancer, with the current focus placed on pharmaceutical intervention to restrict androgen synthesis selectively, an endeavor that could benefit from the enhanced understanding of enzymatic mechanisms that derives from characterization of key reaction intermediates. The multifunctional cytochrome P450 17A1 (CYP17A1) first catalyzes the typical hydroxylation of its primary substrate, pregnenolone (PREG) and then also orchestrates a remarkable C17-C20 bond cleavage (lyase) reaction, converting the 17-hydroxypregnenolone initial product to dehydroepiandrosterone, a process representing the first committed step in the biosynthesis of androgens. Now, we report the capture and structural characterization of intermediates produced during this lyase step: an initial peroxo-anion intermediate, poised for nucleophilic attack on the C20 position by a substrate-associated H-bond, and the crucial ferric peroxo-hemiacetal intermediate that precedes carbon-carbon (C-C) bond cleavage. These studies provide a rare glimpse at the actual structural determinants of a chemical transformation that carries profound physiological consequences.

AB - Ablation of androgen production through surgery is one strategy against prostate cancer, with the current focus placed on pharmaceutical intervention to restrict androgen synthesis selectively, an endeavor that could benefit from the enhanced understanding of enzymatic mechanisms that derives from characterization of key reaction intermediates. The multifunctional cytochrome P450 17A1 (CYP17A1) first catalyzes the typical hydroxylation of its primary substrate, pregnenolone (PREG) and then also orchestrates a remarkable C17-C20 bond cleavage (lyase) reaction, converting the 17-hydroxypregnenolone initial product to dehydroepiandrosterone, a process representing the first committed step in the biosynthesis of androgens. Now, we report the capture and structural characterization of intermediates produced during this lyase step: an initial peroxo-anion intermediate, poised for nucleophilic attack on the C20 position by a substrate-associated H-bond, and the crucial ferric peroxo-hemiacetal intermediate that precedes carbon-carbon (C-C) bond cleavage. These studies provide a rare glimpse at the actual structural determinants of a chemical transformation that carries profound physiological consequences.

KW - Cytochrome P450

KW - Nanodiscs

KW - Peroxo-hemiacetal

KW - Resonance Raman spectroscopy

KW - Steroids

UR - http://www.scopus.com/inward/record.url?scp=84952674074&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84952674074&partnerID=8YFLogxK

U2 - 10.1073/pnas.1519376113

DO - 10.1073/pnas.1519376113

M3 - Article

C2 - 26668369

AN - SCOPUS:84952674074

VL - 112

SP - 15856

EP - 15861

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 - 52

ER -