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Titel: Resurrection and characterization of ancestral CYP11A1 enzymes
VerfasserIn: Hartz, Philip
Strohmaier, Silja J.
El-Gayar, Basma M.
Abdulmughni, Ammar
Hutter, Michael C.
Hannemann, Frank
Gillam, Elizabeth M. J.
Bernhardt, Rita
Sprache: Englisch
Titel: The FEBS Journal
Bandnummer: 288
Heft: 22
Seiten: 6510-6527
Verlag/Plattform: Wiley
Erscheinungsjahr: 2021
Freie Schlagwörter: ancestral sequence reconstruction
cholesterol
cytochromes P450
pregnenolone formation
pregnenolone formation
DDC-Sachgruppe: 570 Biowissenschaften, Biologie
Dokumenttyp: Journalartikel / Zeitschriftenartikel
Abstract: Mitochondrial cytochromes P450 presumably originated from a common microsomal P450 ancestor. However, it is still unknown how ancient mitochondrial P450s were able to retain their oxygenase function following relocation to the mitochondrial matrix and later emerged as enzymes specialized for steroid hormone biosynthesis in vertebrates. Here, we used the approach of ancestral sequence reconstruction (ASR) to resurrect ancient CYP11A1 enzymes and characterize their unique biochemical properties. Two ancestral CYP11A1 variants, CYP11A_Mammal_N101 and CYP11A_N1, as well as an extant bovine form were recombinantly expressed and purified to homogeneity. All enzymes showed characteristic P450 spectral properties and were able to convert cholesterol as well as other sterol substrates to pregnenolone, yet with different specificities. The vertebrate CYP11A_N1 ancestor preferred the cholesterol precursor, desmosterol, as substrate suggesting a convergent evolution of early cholesterol metabolism and CYP11A1 enzymes. Both ancestors were able to withstand increased levels of hydrogen peroxide but only the ancestor CYP11A_N1 showed increased thermostability (˜ 25 °C increase in T50 ) compared with the extant CYP11A1. The extraordinary robustness of ancient mitochondrial P450s, as demonstrated for CYP11A_N1, may have allowed them to stay active when presented with poorly compatible electron transfer proteins and resulting harmful ROS in the new environment of the mitochondrial matrix. To the best of our knowledge, this work represents the first study that describes the resurrection of ancient mitochondrial P450 enzymes. The results will help to understand and gain fundamental functional insights into the evolutionary origins of steroid hormone biosynthesis in animals.
DOI der Erstveröffentlichung: 10.1111/febs.16054
URL der Erstveröffentlichung: https://febs.onlinelibrary.wiley.com/doi/10.1111/febs.16054
Link zu diesem Datensatz: urn:nbn:de:bsz:291--ds-400931
hdl:20.500.11880/36096
http://dx.doi.org/10.22028/D291-40093
ISSN: 1742-4658
Datum des Eintrags: 12-Jul-2023
Drittmittel / Förderung: Universitätsgesellschaft des Saarlandes
Bezeichnung des in Beziehung stehenden Objekts: Supporting Information
In Beziehung stehendes Objekt: https://febs.onlinelibrary.wiley.com/action/downloadSupplement?doi=10.1111%2Ffebs.16054&file=febs16054-sup-0001-Supinfo.pdf
Fakultät: NT - Naturwissenschaftlich- Technische Fakultät
Fachrichtung: NT - Biowissenschaften
Professur: NT - Prof. Dr. Volkhard Helms
NT - Prof. Dr. Bruce Morgan
NT - Keiner Professur zugeordnet
Sammlung:SciDok - Der Wissenschaftsserver der Universität des Saarlandes

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