Resurrection and characterization of ancestral CYP11A1 enzymes.

The FEBS Journal
Philip HartzR Bernhardt

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 robus...Continue Reading

References

Sep 15, 1978·The Biochemical Journal·J I MasonG S Boyd
Nov 16, 1992·Biochemical and Biophysical Research Communications·H UhlmannR Bernhardt
Oct 5, 1990·Journal of Molecular Biology·S F AltschulD J Lipman
Mar 1, 1990·Journal of Molecular Graphics·G Vriend
Jun 1, 1989·Archives of Biochemistry and Biophysics·C R BhaskerM R Waterman
Oct 14, 1970·Journal of Molecular Biology·M Mandel, A Higa
May 28, 1982·Molecular and Cellular Biochemistry·J D LambethH Kamin
Oct 1, 1993·European Journal of Biochemistry·R C Tuckey, K J Cameron
Jul 1, 1993·Biological & Pharmaceutical Bulletin·Y SagaraT Horiuchi
Jun 20, 1997·Journal of Molecular Biology·G VogtP Argos
Feb 17, 1999·Annual Review of Entomology·R Feyereisen
Mar 31, 1999·Progress in Biophysics and Molecular Biology·R Jaenicke
Jul 19, 2000·Proteins·A V GrinbergR Bernhardt
Feb 24, 2001·Genome Biology·D Werck-Reichhart, R Feyereisen
Aug 23, 2001·Proceedings of the National Academy of Sciences of the United States of America·N A BakerJ A McCammon
Jun 25, 2004·Nucleic Acids Research·Todd J DolinskyNathan A Baker
Apr 7, 2006·Biochemical and Biophysical Research Communications·Miharu AraseNorio Kagawa
Jun 23, 2006·PLoS Computational Biology·Paul D WilliamsRichard A Goldstein
Sep 19, 2006·Biochimica Et Biophysica Acta·Frank HannemannRita Bernhardt
Jan 20, 2009·Bioinformatics·Andrew M WaterhouseGeoffrey J Barton
Dec 3, 2009·Human Genomics·David R Nelson
Jan 1, 2009·Journal of Cheminformatics·Tobias Lippert, Matthias Rarey
Jun 12, 2010·Biochimica Et Biophysica Acta·Kerstin Maria EwenRita Bernhardt
Jul 14, 2010·Biochimica Et Biophysica Acta·B Crešnar, S Petrič
Oct 14, 2010·Archives of Biochemistry and Biophysics·Kip P ConnerWilliam M Atkins
Nov 9, 2010·Applied Microbiology and Biotechnology·Anett SchallmeyDick B Janssen
Dec 3, 2010·Methods in Molecular Biology·Norio Kagawa
Dec 8, 2010·Bioinformatics·Pascal BenkertTorsten Schwede
Dec 17, 2010·The Journal of Biological Chemistry·Natalia MastIrina A Pikuleva
Dec 21, 2010·Archives of Biochemistry and Biophysics·Abhinav LuthraStephen G Sligar
Jun 4, 2011·Proceedings of the National Academy of Sciences of the United States of America·Natallia StrushkevichHee-Won Park

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.