Small angle scattering reveals the orientation of cytochrome P450 19A1 in lipoprotein nanodiscs

John C. Hackett; Susan Krueger; Volker S. Urban; Francisco Zárate-Pérez

doi:10.1016/j.jinorgbio.2024.112579

Small angle scattering reveals the orientation of cytochrome P450 19A1 in lipoprotein nanodiscs

John C. Hackett
, Susan Krueger
, Volker S. Urban
, Francisco Zárate-Pérez

Neutron Scattering Division

Research output: Contribution to journal › Article › peer-review

Abstract

Human aromatase (CYP19A1), the cytochrome P450 enzyme responsible for conversion of androgens to estrogens, was incorporated into lipoprotein nanodiscs (NDs) and interrogated by small angle X-ray and neutron scattering (SAXS/SANS). CYP19A1 was associated with the surface and centered at the edge of the long axis of the ND membrane. In the absence of the N-terminal anchor, the amphipathic A'- and G'-helices were predominately buried in the lipid head groups, with the possibly that their hydrophobic side chains protrude into the hydrophobic, aliphatic tails. The prediction is like that for CYP3A4 based on SAXS employing a similar modeling approach. The orientation of CYP19A1 in a ND is consistent with our previous predictions based on molecular dynamics simulations and lends additional credibility to the notion that CYP19A1 captures substrates from the membrane.

Original language	English
Article number	112579
Journal	Journal of Inorganic Biochemistry
Volume	257
DOIs	https://doi.org/10.1016/j.jinorgbio.2024.112579
State	Published - Aug 2024

Funding

This work was supported by National Institutes of Health grant R01GM135414 awarded to J. C. H. Neutron scattering research conducted using the Bio-SANS instrument, a DOE Office of Science, Office of Biological and Environmental Research resource (FWP ERKP291), used resources at the High-Flux Isotope Reactor, a DOE Office of Science, Scientific User Facility operated by the Oak Ridge National Laboratory. This work was also conducted at the Advanced Light Source (ALS), a national user facility operated by Lawrence Berkeley National Laboratory on behalf of the Department of Energy, Office of Basic Energy Sciences, through the Integrated Diffraction Analysis Technologies (IDAT) program, supported by DOE Office of Biological and Environmental Research. Additional support comes from the National Institute of Health project ALS-ENABLE (P30 GM124169) and a High-End Instrumentation Grant S10OD018483.

Keywords

Cytochrome P450
Membrane protein
Nanodisc
SANS
SAXS

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10.1016/j.jinorgbio.2024.112579

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title = "Small angle scattering reveals the orientation of cytochrome P450 19A1 in lipoprotein nanodiscs",

abstract = "Human aromatase (CYP19A1), the cytochrome P450 enzyme responsible for conversion of androgens to estrogens, was incorporated into lipoprotein nanodiscs (NDs) and interrogated by small angle X-ray and neutron scattering (SAXS/SANS). CYP19A1 was associated with the surface and centered at the edge of the long axis of the ND membrane. In the absence of the N-terminal anchor, the amphipathic A'- and G'-helices were predominately buried in the lipid head groups, with the possibly that their hydrophobic side chains protrude into the hydrophobic, aliphatic tails. The prediction is like that for CYP3A4 based on SAXS employing a similar modeling approach. The orientation of CYP19A1 in a ND is consistent with our previous predictions based on molecular dynamics simulations and lends additional credibility to the notion that CYP19A1 captures substrates from the membrane.",

keywords = "Cytochrome P450, Membrane protein, Nanodisc, SANS, SAXS",

author = "Hackett, \{John C.\} and Susan Krueger and Urban, \{Volker S.\} and Francisco Z{\'a}rate-P{\'e}rez",

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doi = "10.1016/j.jinorgbio.2024.112579",

language = "English",

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T1 - Small angle scattering reveals the orientation of cytochrome P450 19A1 in lipoprotein nanodiscs

AU - Hackett, John C.

AU - Krueger, Susan

AU - Urban, Volker S.

AU - Zárate-Pérez, Francisco

PY - 2024/8

Y1 - 2024/8

N2 - Human aromatase (CYP19A1), the cytochrome P450 enzyme responsible for conversion of androgens to estrogens, was incorporated into lipoprotein nanodiscs (NDs) and interrogated by small angle X-ray and neutron scattering (SAXS/SANS). CYP19A1 was associated with the surface and centered at the edge of the long axis of the ND membrane. In the absence of the N-terminal anchor, the amphipathic A'- and G'-helices were predominately buried in the lipid head groups, with the possibly that their hydrophobic side chains protrude into the hydrophobic, aliphatic tails. The prediction is like that for CYP3A4 based on SAXS employing a similar modeling approach. The orientation of CYP19A1 in a ND is consistent with our previous predictions based on molecular dynamics simulations and lends additional credibility to the notion that CYP19A1 captures substrates from the membrane.

AB - Human aromatase (CYP19A1), the cytochrome P450 enzyme responsible for conversion of androgens to estrogens, was incorporated into lipoprotein nanodiscs (NDs) and interrogated by small angle X-ray and neutron scattering (SAXS/SANS). CYP19A1 was associated with the surface and centered at the edge of the long axis of the ND membrane. In the absence of the N-terminal anchor, the amphipathic A'- and G'-helices were predominately buried in the lipid head groups, with the possibly that their hydrophobic side chains protrude into the hydrophobic, aliphatic tails. The prediction is like that for CYP3A4 based on SAXS employing a similar modeling approach. The orientation of CYP19A1 in a ND is consistent with our previous predictions based on molecular dynamics simulations and lends additional credibility to the notion that CYP19A1 captures substrates from the membrane.

KW - Cytochrome P450

KW - Membrane protein

KW - Nanodisc

KW - SANS

KW - SAXS

UR - https://www.scopus.com/pages/publications/85192063390

U2 - 10.1016/j.jinorgbio.2024.112579

DO - 10.1016/j.jinorgbio.2024.112579

M3 - Article

C2 - 38703512

AN - SCOPUS:85192063390

SN - 0162-0134

VL - 257

JO - Journal of Inorganic Biochemistry

JF - Journal of Inorganic Biochemistry

M1 - 112579

ER -

Small angle scattering reveals the orientation of cytochrome P450 19A1 in lipoprotein nanodiscs

Abstract

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Keywords

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