Elucidation of stability determinants of cold-adapted monomeric isocitrate dehydrogenase from a psychrophilic bacterium, Colwellia maris, by construction of chimeric enzymes

Microbiology
Seiya WatanabeYasuhiro Takada

Abstract

To elucidate determinants of differences in thermostability between mesophilic and psychrophilic monomeric isocitrate dehydrogenases (IDHs) from Azotobacter vinelandii (AvIDH) and Colwellia maris (CmIDH), respectively, chimeric enzymes derived from the two IDHs were constructed based on the recently resolved three-dimensional structure of AvIDH, and several characteristics of the two wild-type and six chimeric IDHs were examined. These characteristics were then compared with those of dimeric IDH from Escherichia coli (EcIDH). All recombinant enzymes with a (His)(6)-tag attached to the N-terminal were overexpressed in the E. coli cells and purified by Ni(2+)-affinity chromatography. The catalytic activity (k(cat)) and catalytic efficiency (k(cat)/K(m)) of the wild-type AvIDH and CmIDH were higher than those of EcIDH, implying that an improved catalytic rate more than compensates for the loss of a catalytic site in the former two IDHs due to monomerization. Analyses of the thermostability and kinetic parameters of the chimeric enzymes indicated that region 2, corresponding to domain II, and particularly region 3 located in the C-terminal part of domain I, are involved in the thermolability of CmIDH, and that the corresponding two...Continue Reading

References

Nov 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·J H HurleyR M Stroud
Oct 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·B W MatthewsW J Becktel
Dec 16, 1970·Biochimica Et Biophysica Acta·C R Barrera, P Jurtshuk
Apr 1, 1997·Proceedings of the National Academy of Sciences of the United States of America·Antony M Dean, G B Golding
Dec 10, 1997·Biochimica Et Biophysica Acta·Charles GerdayGeorges Feller
Sep 16, 1998·Proceedings of the National Academy of Sciences of the United States of America·P A Fields, G N Somero
Nov 26, 1998·International Journal of Systematic Bacteriology·I YumotoH Okuyama
Mar 29, 2000·Journal of Molecular Biology·K MiyazakiF H Arnold
Feb 24, 2001·Archives of Biochemistry and Biophysics·R Chen, H Yang
Feb 24, 2001·Protein Science : a Publication of the Protein Society·R Chen, S S Jeong
Jun 26, 2001·Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology·P A Fields
Jun 27, 2002·Journal of Molecular Biology·Maaike de BackerEdward Hough
Nov 5, 2002·The Journal of Biological Chemistry·Suren AghajanianPaul C Engel

Citations

Jun 16, 2010·Archives of Microbiology·Shougo MatsuoYasuhiro Takada
Jan 19, 2006·Extremophiles : Life Under Extreme Conditions·Shinsuke MakiYasuhiro Takada
Feb 6, 2009·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Hege Lynum PedersenIngar Leiros
Jan 22, 2008·Applied and Environmental Microbiology·Debora Frigi Rodrigues, James M Tiedje
Jun 8, 2006·Annual Review of Biochemistry·Khawar Sohail Siddiqui, Ricardo Cavicchioli
Oct 31, 2012·International Journal of Molecular Sciences·Caroline Struvay, Georges Feller
Sep 27, 2016·Frontiers in Microbiology·Margarita SantiagoLoreto P Parra
Jan 1, 2013·Biology·Charles Gerday
Apr 28, 2017·Extremophiles : Life Under Extreme Conditions·Ryo HirotaYasuhiro Takada
Mar 13, 2018·FEMS Microbiology Ecology·Isabelle Raymond-BouchardLyle G Whyte

Related Concepts

Adaptation, Physiological
Cold Temperature
DNA, Bacterial
Enzyme Stability
Alkalescens-Dispar Group
Isocitrate Dehydrogenase-I
Protein Conformation
Chimeric Proteins, Recombinant
Trypure
Azotobacter vinelandii

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.

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.

Systemic Juvenile Idiopathic Arthritis

Systemic juvenile idiopathic arthritis is a rare rheumatic disease that affects children. Symptoms include joint pain, but also fevers and skin rashes. Here is the latest on this disease.

Chromatin Regulation and Circadian Clocks

The circadian clock plays an important role in regulating transcriptional dynamics through changes in chromatin folding and remodelling. Discover the latest research on Chromatin Regulation and Circadian Clocks here.

Central Pontine Myelinolysis

Central Pontine Myelinolysis is a neurologic disorder caused most frequently by rapid correction of hyponatremia and is characterized by demyelination that affects the central portion of the base of the pons. Here is the latest research on this disease.

Myocardial Stunning

Myocardial stunning is a mechanical dysfunction that persists after reperfusion of previously ischemic tissue in the absence of irreversible damage including myocardial necrosis. Here is the latest research.

Pontocerebellar Hypoplasia

Pontocerebellar hypoplasias are a group of neurodegenerative autosomal recessive disorders with prenatal onset, atrophy or hypoplasia of the cerebellum, hypoplasia of the ventral pons, microcephaly, variable neocortical atrophy and severe mental and motor impairments. Here is the latest research on pontocerebellar hypoplasia.

Cell Atlas Along the Gut-Brain Axis

Profiling cells along the gut-brain axis at the single cell level will provide unique information for each cell type, a three-dimensional map of how cell types work together to form tissues, and insights into how changes in the map underlie health and disease of the GI system and its crosstalk with the brain. Disocver the latest research on single cell analysis of the gut-brain axis here.

Chronic Traumatic Encephalopathy

Chronic Traumatic Encephalopathy (CTE) is a progressive degenerative disease that occurs in individuals that suffer repetitive brain trauma. Discover the latest research on traumatic encephalopathy here.