Kinetics and thermodynamics of the thermal inactivation and chaperone assisted folding of zebrafish dihydrofolate reductase

Thapliyal, Charu; Jain, Neha; Rashid, Naira; Chaudhuri, Pratima

doi:10.1016/j.abb.2017.11.006

Cited by 4 publications

(2 citation statements)

References 40 publications

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“…Recently, dhfr transgenic lines have become available to identify expression dynamics [ 93 ]. Zebrafish dhfr has also been used to investigate chaperone assisted refolding [ 94 ]. Furthermore, a light-inducible drug for timed inhibition of DHFR is available and could be used for further investigations in zebrafish [ 95 ].…”

Section: Metabolism Of Vitamins and Co-factorsmentioning

confidence: 99%

A Great Catch for Investigating Inborn Errors of Metabolism—Insights Obtained from Zebrafish

Breuer

Patten

2020

Biomolecules

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Inborn errors of metabolism cause abnormal synthesis, recycling, or breakdown of amino acids, neurotransmitters, and other various metabolites. This aberrant homeostasis commonly causes the accumulation of toxic compounds or depletion of vital metabolites, which has detrimental consequences for the patients. Efficient and rapid intervention is often key to survival. Therefore, it requires useful animal models to understand the pathomechanisms and identify promising therapeutic drug targets. Zebrafish are an effective tool to investigate developmental mechanisms and understanding the pathophysiology of disorders. In the past decades, zebrafish have proven their efficiency for studying genetic disorders owing to the high degree of conservation between human and zebrafish genes. Subsequently, several rare inherited metabolic disorders have been successfully investigated in zebrafish revealing underlying mechanisms and identifying novel therapeutic targets, including methylmalonic acidemia, Gaucher’s disease, maple urine disorder, hyperammonemia, TRAPPC11-CDGs, and others. This review summarizes the recent impact zebrafish have made in the field of inborn errors of metabolism.

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Section: Metabolism Of Vitamins and Co-factorsmentioning

confidence: 99%

A Great Catch for Investigating Inborn Errors of Metabolism—Insights Obtained from Zebrafish

Breuer

Patten

2020

Biomolecules

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“…For example, thermal denaturation has been used to analyze thermodynamics of hybridization on solid supports including effects of chain length [1][2][3], probe associations [4,5] and surface density [6], probe-surface interactions [7,8], salt concentration [9][10][11][12][13][14], and how such systems perform in commercial technologies [15]. Similar needs arise for immobilized protein films such as antibodies or enzymes [16][17][18], where the impact of the surface environment on stability or activity of the protein could be correlated with determination of its denaturation transition and unfolding. Such applications require immobilization chemistries capable of performing at temperatures approaching physical limits, like the boiling point of water.…”

Section: Introductionmentioning

confidence: 99%

Electrochemical Analysis of Ultrathin Polythiolsiloxane Films for Surface Biomodification

Chiang¹,

Levicky²

2018

International Journal of Electrochemistry

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The ability of different crosslinkers to crosslink nanometer thick films of the polymer poly(mercaptopropyl)methylsiloxane (PMPMS), thus stabilizing these films on solid supports, was investigated. The four crosslinkers included 1,11-bismaleimidotriethyleneglycol (BM(PEG)3), tris-(2-maleimidoethyl)amine (TMEA), bismaleimidohexane (BMH), and 1,1′-(methylenedi-4,1-phenylene) bismaleimide (BMDPM). PMPMS films treated with the four crosslinkers were compared in the effectiveness of achieved crosslinking, continuity and stability of the films to rearrangement at elevated temperatures, and modification with single-stranded DNA. The results of electrochemical analyses show that more hydrophilic crosslinkers had difficulty reacting fully with PMPMS thiols, even in these nanometer thin layers. This observation highlights the critical importance of selecting crosslinkers that are chemically compatible. Optimal selection of crosslinker yielded films in which the polymer film was largely incapable of rearranging, even at elevated temperatures, yielding reproducible and stable layers. These results validate use of these supports for applications such as monitoring thermal denaturation of immobilized DNA duplexes.

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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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Kinetics and thermodynamics of the thermal inactivation and chaperone assisted folding of zebrafish dihydrofolate reductase

Cited by 4 publications

References 40 publications

A Great Catch for Investigating Inborn Errors of Metabolism—Insights Obtained from Zebrafish

A Great Catch for Investigating Inborn Errors of Metabolism—Insights Obtained from Zebrafish

Electrochemical Analysis of Ultrathin Polythiolsiloxane Films for Surface Biomodification

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

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