Investigation of membrane disruption in the reaction catalyzed by cholesterol oxidase

Biochemistry
K B GhoshroyNicole S Sampson

Abstract

Dye leakage experiments were undertaken to investigate the membrane disruption properties of cholesterol oxidase. Inspection of the X-ray crystal structures of cholesterol oxidase suggested that an active-site "lid" opens in order to bind substrate [Li, J., Vrielink, A., Brick, P., & Blow, D. M. (1993) Biochemistry 32, 11507-11515]. We tested whether the interaction of the putative active-site lid with the membrane was sufficiently disruptive of the membrane structure to cause leakage or lysis of the cell membrane. Vesicles (100 nm) composed of egg phosphatidylcholine, 2-palmitoyl-3-oleoyl-1-sn-phosphatidylethanolamine, and 2-palmitoyl-3-oleoyl-1-sn-phosphatidylcholine were used in this study to mimic biomembranes. To separate the effects of membrane binding from conversion of cholesterol to cholest-4-en-3-one, the active-site mutant E361Q was utilized. In the reaction catalyzed by E361Q, isomerization of the cholest-5-en-3-one intermediate is suppressed and cholest-5-en-3-one is the major product isolated. Furthermore, E361Q produces cholest-5-en-3-one 20-fold more slowly than wild type produces cholest-4-en-3-one from cholesterol. Wild-type and E361Q cholesterol oxidases bind to vesicles with an apparent K(D) of approximately...Continue Reading

References

Jan 1, 1989·Annual Review of Biophysics and Biophysical Chemistry·T M Jovin, D J Arndt-Jovin
Jun 24, 1987·Biochimica Et Biophysica Acta·M C PhillipsG H Rothblat

❮ Previous
Next ❯

Citations

Aug 2, 2007·Bioscience Reports·Gerald Gimpl, Katja Gehrig-Burger
May 24, 2000·The Journal of Steroid Biochemistry and Molecular Biology·J MacLachlanC J Brooks
Sep 29, 2001·Biomolecular Engineering·P Walde, S Ichikawa
Jul 18, 2008·Molecular BioSystems·Jesús F Aparicio, Juan F Martín
Sep 21, 2002·Journal of Veterinary Medicine. B, Infectious Diseases and Veterinary Public Health·H FuhrmannT Gück
May 9, 2012·Proceedings of the National Academy of Sciences of the United States of America·Hubert M PiwonskiGilad Haran
Aug 30, 2005·The Journal of Clinical Investigation·Luigi PuglielliAshley I Bush
Apr 22, 2008·Progress in Lipid Research·Yvonne Lange, Theodore L Steck
Feb 9, 2012·Microbial Biotechnology·J L GarcíaB Galán
Oct 22, 2009·The FEBS Journal·Joseph Kreit, Nicole S Sampson
Mar 24, 2007·Chemistry & Biology·Natasha M Nesbitt, Nicole S Sampson
Jul 1, 2016·Journal of Pharmaceutical and Biomedical Analysis·Atsushi ShojiMasao Sugawara
Jul 20, 2016·Biochimica Et Biophysica Acta·Elisa La BauveMichael S Kent
Sep 17, 2003·Accounts of Chemical Research·Nicole S Sampson, Alice Vrielink

❮ 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.