Nanodiscs in the studies of membrane-bound cytochrome P450 enzymes

A. Luthra, M. Gregory, Y. V. Grinkova, I. G. Denisov, Stephen Sligar

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Cytochromes P450 from eukaryotes and their native redox partners cytochrome P450 reductases both belong to the class of monotopic membrane proteins containing one transmembrane anchor. Incorporation into the lipid bilayer signi ficantly affects their equilibrium and kinetic properties and plays an important role in their interactions. We describe here the detailed protocols developed in our group for the functional self-assembly of mammalian cytochromes P450 and cytochrome P450 reductases into Nanodiscs with controlled lipid composition. The resulting preparations are fully functional, homogeneous in size, composition and oligomerization state of the heme enzyme, and show an improved stability with respect to P420 formation. We provide a brief overview of applications of Nanodisc technology to the biophysical and biochemical mechanistic studies of cytochromes P450 involved in steroidogenesis, and of the most abundant xenobiotic-metabolizing human cytochrome P450 CYP3A4.

Original languageEnglish (US)
Title of host publicationCytochrome P450 Protocols
PublisherHumana Press Inc.
Pages115-127
Number of pages13
ISBN (Print)9781627033206
DOIs
StatePublished - 2013

Publication series

NameMethods in Molecular Biology
Volume987
ISSN (Print)1064-3745

Fingerprint

Cytochrome P-450 Enzyme System
NADPH-Ferrihemoprotein Reductase
Membranes
Cytochrome P-450 CYP3A
Lipid Bilayers
Xenobiotics
Eukaryota
Heme
Oxidation-Reduction
Membrane Proteins
Technology
Lipids
Enzymes

Keywords

  • CYP17
  • CYP19A1
  • CYP3A4
  • Cytochrome P450
  • Eukaryotic P450
  • Membrane mimetics
  • Nanodiscs

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Luthra, A., Gregory, M., Grinkova, Y. V., Denisov, I. G., & Sligar, S. (2013). Nanodiscs in the studies of membrane-bound cytochrome P450 enzymes. In Cytochrome P450 Protocols (pp. 115-127). (Methods in Molecular Biology; Vol. 987). Humana Press Inc.. https://doi.org/10.1007/978-1-62703-321-3_10

Nanodiscs in the studies of membrane-bound cytochrome P450 enzymes. / Luthra, A.; Gregory, M.; Grinkova, Y. V.; Denisov, I. G.; Sligar, Stephen.

Cytochrome P450 Protocols. Humana Press Inc., 2013. p. 115-127 (Methods in Molecular Biology; Vol. 987).

Research output: Chapter in Book/Report/Conference proceedingChapter

Luthra, A, Gregory, M, Grinkova, YV, Denisov, IG & Sligar, S 2013, Nanodiscs in the studies of membrane-bound cytochrome P450 enzymes. in Cytochrome P450 Protocols. Methods in Molecular Biology, vol. 987, Humana Press Inc., pp. 115-127. https://doi.org/10.1007/978-1-62703-321-3_10
Luthra A, Gregory M, Grinkova YV, Denisov IG, Sligar S. Nanodiscs in the studies of membrane-bound cytochrome P450 enzymes. In Cytochrome P450 Protocols. Humana Press Inc. 2013. p. 115-127. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-62703-321-3_10
Luthra, A. ; Gregory, M. ; Grinkova, Y. V. ; Denisov, I. G. ; Sligar, Stephen. / Nanodiscs in the studies of membrane-bound cytochrome P450 enzymes. Cytochrome P450 Protocols. Humana Press Inc., 2013. pp. 115-127 (Methods in Molecular Biology).
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