Computational site-directed mutagenesis studies of the role of the hydrophobic triad on substrate binding in cholesterol oxidase

Proteins
Laith Hisham HarbRicardo L Mancera

Abstract

Cholesterol oxidase (ChOx) is a flavoenzyme that oxidizes and isomerizes cholesterol (CHL) to form cholest-4-en-3-one. Molecular docking and molecular dynamics simulations were conducted to predict the binding interactions of CHL in the active site. Several key interactions (E361-CHL, N485-FAD, and H447-CHL) were identified and which are likely to determine the correct positioning of CHL relative to flavin-adenine dinucleotide (FAD). Binding of CHL also induced changes in key residues of the active site leading to the closure of the oxygen channel. A group of residues, Y107, F444, and Y446, known as the hydrophobic triad, are believed to affect the binding of CHL in the active site. Computational site-directed mutagenesis of these residues revealed that their mutation affects the conformations of key residues in the active site, leading to non-optimal binding of CHL and to changes in the structure of the oxygen channel, all of which are likely to reduce the catalytic efficiency of ChOx. Proteins 2017; 85:1645-1655. © 2017 Wiley Periodicals, Inc.

References

Jan 17, 1995·Biochemical and Biophysical Research Communications·I J Kass, N S Sampson
Jan 5, 1999·Bioorganic & Medicinal Chemistry Letters·I J Kass, N S Sampson
Feb 22, 2002·Acta Crystallographica. Section D, Biological Crystallography·Thomas Oldfield
Sep 17, 2003·Accounts of Chemical Research·Nicole S Sampson, Alice Vrielink
Mar 19, 2004·Journal of Medicinal Chemistry·Richard A FriesnerPeter S Shenkin
Apr 11, 2006·Nature Chemical Biology·Artem Y LyubimovAlice Vrielink
Jul 22, 2006·The Biochemical Journal·Louis LimAlice Vrielink
Nov 22, 2007·Protein Science : a Publication of the Protein Society·Artem Y LyubimovAlice Vrielink
Oct 22, 2009·The FEBS Journal·Joseph Kreit, Nicole S Sampson
Oct 22, 2009·The FEBS Journal·Alice Vrielink, Sandro Ghisla
Mar 10, 2010·Sub-cellular Biochemistry·Alice Vrielink
Apr 23, 2010·BMC Biotechnology·Federica VolontèLuciano Piubelli
Oct 12, 2012·Natural Product Reports·Christopher T Walsh, Timothy A Wencewicz
May 8, 2014·Journal of Chemical Theory and Computation·Callum J DicksonRoss C Walker
May 13, 2014·Protein Engineering, Design & Selection : PEDS·Noeris K SalamDavid A Pearlman
Dec 6, 2014·Acta Crystallographica. Section D, Biological Crystallography·Emily GoldenAlice Vrielink
Nov 18, 2015·Journal of Chemical Theory and Computation·James A MaierCarlos Simmerling

❮ Previous
Next ❯

Citations


❮ 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.