Disruption of an ionic network leads to accelerated thermal denaturation of D-glyceraldehyde-3-phosphate dehydrogenase from the hyperthermophilic bacterium Thermotoga maritima

Journal of Molecular Biology
G PappenbergerR Jaenicke

Abstract

The role of an ionic network of four charged amino acid side-chains in the thermostability of the enzyme D-glyceraldehyde-3-phosphate dehydrogenase from the hyperthermophilic bacterium Thermotoga maritima (TmGAPDH) has been assessed by site-directed mutagenesis, replacing the central residue of the ionic network, arginine 20, by either alanine (R20A) or asparagine (R20N). The purified mutant enzymes display no differences to the wild-type enzyme regarding spectroscopic properties and enzymatic activity. However, denaturation kinetics reveal that the resistance towards thermal denaturation is strongly diminished in the mutant enzymes. This is reflected by a decrease in free energy of activation for thermal unfolding of about 4 kJ/mol at 100 degrees C and a shift of temperature of half denaturation after one hour incubation from 96 to 89 degrees C for both mutant enzymes. Due to a large decrease in activation enthalpy, the effects of the mutations are temperature dependent and become even more significant at the physiological temperature of Thermotoga maritima (approximately 80 degrees C). The importance of the arginine 20 side-chain for kinetic thermal stability is plausible in the light of its key role in the ionic network and ...Continue Reading

Citations

Nov 25, 1998·Protein Science : a Publication of the Protein Society·E J SpekN R Kallenbach
Jun 7, 2012·Chemical Research in Toxicology·Shanen M SherrerZucai Suo
Apr 7, 1999·The Biochemical Journal·E PedoneS Bartolucci
Mar 10, 2001·Microbiology and Molecular Biology Reviews : MMBR·C Vieille, G J Zeikus
May 30, 1998·Proceedings of the National Academy of Sciences of the United States of America·R C WadeV Lounnas
Dec 6, 2001·Journal of Molecular Biology·D DertingerO C Uhlenbeck
Jun 26, 2003·Protein Science : a Publication of the Protein Society·Kam-Bo WongMark Bycroft
Mar 8, 2007·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Charles K LeeMichelle E Peterson
Apr 28, 2001·The Journal of Biological Chemistry·S D'AmicoG Feller
Dec 11, 2002·European Journal of Biochemistry·Graeme S BellGarry L Taylor
Nov 30, 2018·Frontiers in Microbiology·Rubén DíazLeonardo David Herrera-Zúñiga
Aug 13, 2015·Cellular and Molecular Life Sciences : CMLS·Francesco ItriAngela Arciello
Aug 31, 2013·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Mohammad Wadud BhuiyaBernard Andrew Brown
Mar 31, 1999·Progress in Biophysics and Molecular Biology·R Jaenicke
Mar 21, 2021·Diagnostic Microbiology and Infectious Disease·Enoch AninagyeiDesmond Omane Acheampong
Nov 18, 2000·Structure·T KajanderA Goldman
Jan 11, 2001·Biochimica Et Biophysica Acta·P P SheridanJ E Brenchley
Sep 19, 2017·International Journal of Biological Macromolecules·Mehrnaz Azami-MovahedMohsen Nemat-Gorgani
Sep 25, 1998·Trends in Microbiology·M J Danson, D W Hough
Jun 27, 2000·Journal of Biotechnology·R Jaenicke
Apr 15, 2003·Journal of Molecular Biology·Tony CollinsGeorges Feller

❮ 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.