Refined method to study the posttranslational regulation of alternative oxidases from Arabidopsis thaliana in vitro

Physiologia Plantarum
Jennifer SelinskiRenate Scheibe

Abstract

In isolated membranes, posttranslational regulation of quinol oxidase activities can only be determined simultaneously for all oxidases - quinol oxidases as well as cytochrome c oxidases - because of their identical localization. In this study, a refined method to determine the specific activity of a single quinol oxidase is exemplarily described for the alternative oxidase (AOX) isoform AOX1A from Arabidopsis thaliana and its corresponding mutants, using the respiratory chain of an Escherichia coli cytochrome bo and bd-I oxidase double mutant as a source to provide electrons necessary for O2 reduction via quinol oxidases. A highly sensitive and reproducible experimental set-up with prolonged linear time intervals of up to 60 s is presented, which enables the determination of constant activity rates in E. coli membrane vesicles enriched in the quinol oxidase of interest by heterologous expression, using a Clark-type oxygen electrode to continuously follow O2 consumption. For the calculation of specific quinol oxidase activity, activity rates were correlated with quantitative signal intensity determinations of AOX1A present in a membrane-bound state by immunoblot analyses, simultaneously enabling normalization of specific activi...Continue Reading

References

Nov 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·A M Kumar, D Söll
Jun 1, 1992·Trends in Biochemical Sciences·D JahnD Söll
Apr 23, 1991·Biochemistry·A PuustinenM Wikström
Jan 2, 1986·European Journal of Biochemistry·C FlintaG von Heijne
Aug 1, 1968·Journal of Bacteriology·A SăsărmanA Dumitrescu
Jun 16, 1995·Journal of Molecular Biology·J M ThornK J Edwards
Jul 1, 1994·Plant Physiology·L McIntosh
Nov 1, 1996·European Journal of Biochemistry·M G Claros, P Vincens
Aug 22, 1997·Biochimica Et Biophysica Acta·S Jünemann
Jun 1, 2000·Proceedings of the National Academy of Sciences of the United States of America·K A Datsenko, B L Wanner
Sep 27, 2000·Biochimica Et Biophysica Acta·C R Lancaster, A Kröger
May 30, 2002·Biochimica Et Biophysica Acta·Ann L UmbachJames N Siedow
Apr 1, 1964·Biochemistry·W W CLELAND
Sep 30, 2003·Biotechnology Letters·Antonio Villaverde, M Mar Carrió
May 12, 2004·Current Opinion in Plant Biology·Alisdair R FernieLee J Sweetlove
Sep 25, 2004·Acta Crystallographica. Section D, Biological Crystallography·Gerlind SulzenbacherChristian Cambillau
Feb 7, 2006·Biochimica Et Biophysica Acta·Ann L UmbachJames N Siedow
May 5, 2006·Journal of Bacteriology·Eric V Patridge, James G Ferry
Jan 5, 2008·Annual Review of Biochemistry·James A Imlay
Sep 13, 2008·The Journal of Biological Chemistry·Monika G DüserMichael Börsch
Oct 28, 2008·Applied and Environmental Microbiology·Vasiliy A PortnoyBernhard Ø Palsson
Feb 3, 2009·Journal of Bacteriology·Britta BallhausenGabriele Deckers-Hebestreit
May 28, 2009·Physiologia Plantarum·Olivier Van AkenJames Whelan
Sep 29, 2009·Physiologia Plantarum·Alexios N PolidorosBirgit Arnholdt-Schmitt
Feb 22, 2011·Annual Review of Plant Biology·A Harvey MillarDavid A Day
May 14, 2011·Biotechnology Journal·Gustavo SalinasCecilia Fernández
Jul 16, 2011·Biochimica Et Biophysica Acta·Vitaliy B BorisovMichael I Verkhovsky
Aug 9, 2011·Nature·Rouslan G Efremov, Leonid A Sazanov
Oct 12, 2011·Proceedings of the National Academy of Sciences of the United States of America·Vitaliy B BorisovMichael I Verkhovsky
Jul 31, 2012·Applied and Environmental Microbiology·Poonam SharmaMartijn Bekker

❮ Previous
Next ❯

Citations

Dec 5, 2018·The Journal of Biological Chemistry·Yuval KayeArthur R Grossman
Sep 6, 2019·G3 : Genes - Genomes - Genetics·Hongli YangJing Liu

❮ 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.