Utilization of positional isotope exchange experiments to evaluate reversibility of ATP hydrolysis catalyzed by Escherichia coli Lon protease

Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire
Jennifer ThomasIrene Lee

Abstract

Lon protease, also known as protease La, is an ATP-dependent serine protease. Despite the presence of a proteolytic Ser-Lys dyad, the enzyme only catalyzes protein degradation in the presence of ATP. Lon possesses an intrinsic ATPase activity that is stimulated by protein and certain peptide substrates. Through sequence alignment and analysis, it is concluded that Lon belongs to the AAA+ protein family. Previous kinetic characterization of the ATPase domain of Escherichia coli Lon protease implicates a half-site reactivity model in which only 50% of the ATP bound to Lon are hydrolyzed to yield ADP; the remaining ATPase sites remain bound with ATP and are considered non-catalytic. In this model, it is implied that ATP hydrolysis is irreversible. To further evaluate the proposed half-site reactivity model, the reversibility of the ATPase activity of E. coli Lon was evaluated by positional isotope exchange experiments. The ATPase reactions were conducted in the 18O-enriched buffer such that the extent of 18O incorporation into inorganic phosphate generated from ATP hydrolysis could be used to evaluate the extent of reversibility in ATP hydrolysis. Collectively, our experimental data reveal that the ATPase reaction catalyzed by E. ...Continue Reading

References

Aug 5, 1991·FEBS Letters· Amerik AYuE V Shimbarevich
Nov 1, 1990·Trends in Biochemical Sciences·M SarasteA Wittinghofer
Aug 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·L Waxman, A L Goldberg
Jan 1, 1980·Methods in Enzymology·D D HackneyP D Boyer
Jan 1, 1994·Methods in Enzymology·A L GoldbergM R Maurizi
Feb 14, 1995·Proceedings of the National Academy of Sciences of the United States of America·G M HeilekH F Noller
Nov 1, 1996·Current Genetics·M Rep, L A Grivell
Apr 1, 1997·Trends in Biochemical Sciences·C K SuzukiG Schatz
Jan 1, 1997·Annual Review of Biochemistry·P D Boyer
Apr 17, 1999·The Journal of Biological Chemistry·D E KimS S Patel
Jul 28, 2001·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·T Ogura, A J Wilkinson
Dec 11, 2003·The Journal of Biological Chemistry·Istvan BotosAlla Gustchina
Mar 24, 2004·Journal of Structural Biology·Istvan BotosAlexander Wlodawer
Dec 21, 2004·European Journal of Biochemistry·Tatyana V RotanovaAlla Gustchina
Aug 3, 2005·Nature Reviews. Molecular Cell Biology·Phyllis I Hanson, Sidney W Whiteheart
Oct 4, 2005·Protein Science : a Publication of the Protein Society·Mi LiAlexander Wlodawer
Dec 13, 2005·Proceedings of the National Academy of Sciences of the United States of America·David D Hackney
Aug 1, 2006·Protein Science : a Publication of the Protein Society·Tatyana V RotanovaAlexander Wlodawer
Aug 22, 2006·The Journal of Biological Chemistry·Adrian O OlivaresEnrique M De La Cruz

❮ Previous
Next ❯

Citations

Jul 27, 2017·Frontiers in Molecular Biosciences·Jennifer FishovitzIrene Lee
Feb 11, 2014·Biochemistry·Michele ScianWilliam M Atkins

❮ 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.