Comparison of physico-chemical aspects between E. coli and human dihydrofolate reductase: an equilibrium unfolding study

Thapliyal, Charu; Jain, Neha; Chaudhuri, Pratima

doi:10.1134/s0006350915030197

Cited by 4 publications

(1 citation statement)

References 19 publications

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“…Inspite of being a pharmacological target, protein chemists has develop great interest in DHFR which becomes a model protein for the research on the structure/functional relationships because of its small size [ 9 ]. The knowledge of the structure/functional relationship is advantageous in the designing of inhibitors of the protein [ 10 ]. zDHFR is a monomer of 190 amino acids with a disulﬁde bond and 3 tryptophan residues and contains many parallel and anti-parallel β sheets which are embedded by α helices.…”

Section: Introductionmentioning

confidence: 99%

A versatile tool in controlling aggregation and Ag nanoparticles assisted in vitro folding of thermally denatured zDHFR

Gupta

Verma

et al. 2020

Biochemistry and Biophysics Reports

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Background Proteins have tendency to form inactive aggregates at higher temperatures due to thermal instability. Maintenance of thermal stability is essential to gain the protein in sufficient quantity and biologically active form during their commercial production. Methods BL21-DE3 Rosetta E. coli cells which contains plasmid pET43.1a vector was used for producing zDHFR protein commercially. The purification of N-terminal Histidine tagged zDHFR was performed by Immobilized Metal Ion chromatography (IMAC). Investigations were performed in existence and non existence of Silver nanoparticles (AgNPs). The inactivation kinetics of zDHFR in existence and non existence of AgNPs were monitored over a range of 40–80 °C as monitored by UV–Visible absorption spectroscopy. Results The protein completely lost its activity at 55 °C. Kinetics of inactivated zDHFR follows first order model in presence and absence of AgNPs. Decrease in rate constant ( k ) values at respective temperatures depicts that AgNPs contribute in the thermostability of the protein. AgNPs also assists in regaining the activity of zDHFR protein. Conclusions AgNPs helps in maintaining thermostability and reducing the aggregation propensity of zDHFR protein. General significance Result explains that AgNPs are recommended as a valuable system in enhancing the industrial production of biologically active zDHFR protein which is an important component in folate cycle and essential for survival of cells and prevents the protein from being aggregated.

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Section: Introductionmentioning

confidence: 99%

A versatile tool in controlling aggregation and Ag nanoparticles assisted in vitro folding of thermally denatured zDHFR

Gupta

Verma

et al. 2020

Biochemistry and Biophysics Reports

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Evolutionarily Related Dihydrofolate Reductases Perform Coequal Functions Yet Show Divergence in Their Trajectories

Rashid

Chattopadhyay

2018

Protein J

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The enzyme dihydrofolate reductase (DHFR) catalyzes NADPH dependent reduction of dihydrofolate to tetrahydrofolate. It plays a crucial role in the DNA synthesis. The investigation of evolution of DHFR generates immense curiosity. It aids in predicting how the enzyme has adapted to the surroundings of various cell types. In spite of great similarity in the structure of E. coli DHFR and human DHFR, their primary sequences are divergent to a great extent, which is evident in variations in the kinetics mechanism of their catalysis. In presence of physiological levels of ligands, they possess distinct kinetics and different rate limiting steps. We have reviewed the process of their unfolding and refolding, their behaviour in denaturing conditions and in presence of various chaperones. Although there is structural similarity between these two homologous enzymes yet they have established distinct mechanisms to accomplish the coequal functions.

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Osmolyte induced enhancement of expression and solubility of human dihydrofolate reductase: An in vivo study

Rashid

Thapliyal

Chaudhuri

2017

International Journal of Biological Macromolecules

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Comparison of physico-chemical aspects between E. coli and human dihydrofolate reductase: an equilibrium unfolding study

Cited by 4 publications

References 19 publications

A versatile tool in controlling aggregation and Ag nanoparticles assisted in vitro folding of thermally denatured zDHFR

A versatile tool in controlling aggregation and Ag nanoparticles assisted in vitro folding of thermally denatured zDHFR

Evolutionarily Related Dihydrofolate Reductases Perform Coequal Functions Yet Show Divergence in Their Trajectories

Osmolyte induced enhancement of expression and solubility of human dihydrofolate reductase: An in vivo study

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