The effect of cysteine-43 mutation on thermostability and kinetic properties of citrate synthase from Thermoplasma acidophilum

Date

1996-07-05

Author

Kocabıyık, Semra
Russel, RJM
Danson, MJ
Hough, DW

Metadata

Show full item record

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Item Usage Stats

338
views

0
downloads

In this study, we have substituted serine-43 by cysteine in the recombinant citrate synthase from a moderately thermophilic Archaeon Thermoplasma acidophilum, for site-specific attachment of labels and have investigated the effects of this mutation on the biochemical properties and thermal stability of the enzyme. Both wild-type and the mutant enzymes were purified to homogenity using affinity chromatography on Matrex Gel Red A. The mutant Thermoplasma citrate synthase is very similar to wild-type citrate synthase in its substrate and co-factor specificities, pH profile and thermal stability. The mutation, however, has decreased the enzyme activity. The newly introduced reactive sulphydryl group could be easily modified by DTNB and labelled with 4-chloro-7-sulphobenzofuran, without loss of any activity. (C) 1996 Academic Press, Inc.

Subject Keywords

Biophysics, Cell Biology, Biochemistry, Molecular Biology

URI

https://hdl.handle.net/11511/49070

Journal

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS

DOI

https://doi.org/10.1006/bbrc.1996.1011

Collections

Department of Biology, Article

Suggestions

OpenMETU
Core

The effect of valine substitution for glycine in the dimer interface of citrate synthase from Thermoplasma acidophilum on stability and activity Kocabıyık, Semra (Elsevier BV, 2000-08-28) To determine the role of hydrophobic interactions in the dimer interface of citrate synthase (CS) from Thermoplasma (Tp) acidophilum in thermostabilization, we have used site-directed mutagenesis to replace Gly 196 by Val on the helix L of the subunit interface. Recombinant wild-type and Gly 196 mutant TpCS enzymes were largely identical in terms of substrate specificities (K-m for oxaloacetate and acetyl CoA). However, the mutation not only reduced catalytic activity (about 10-fold) (i.e., V-max, K-cat and...
Investigating the anticarcinogenic role of salix aegyptiaca l. in colorectal carcinoma Enayat, Shabnam; Banerjee, Sreeparna; Department of Biology (2009) In this study, extracts from bark, leaves and catkins of Salix aegyptiaca L. were investigated for their antioxidant content by 2,2-diphenyl-2-picrylhydrazyl hydrate (DPPH) free radical quenching assay, total phenolic and total flavonoid assays. The highest antioxidant activity (19 ug/ml IC50 for inhibition of DPPH radical activity), total phenolic content (212 mg gallic acid equivalents/g of dried extract) and total flavonoid (479 mg catechin equivalents/g of dried extract) was observed in the ethanolic ex...
Investigating the malleability of RNA aptamers İlgü, Müslüm; Lamm, Monica H.; Nilsen-Hamilton, Marit (Elsevier BV, 2013-09-15) Aptamers are short, single-stranded nucleic acids with structures that frequently change upon ligand binding and are sensitive to the ionic environment. To achieve facile application of aptamers in controlling cellular activities, a better understanding is needed of aptamer ligand binding parameters, structures, intramolecular mobilities and how these structures adapt to different ionic environments with consequent effects on their ligand binding characteristics. Here we discuss the integration of biochemic...
An alternative supporting electrolyte for enzyme immobilization in conducting polymers Kiralp, Senem; Balik, Balam; KARATAŞ, SEVİM; Toppare, Levent Kamil; Gungorb, Atilla (Elsevier BV, 2008-03-01) In this study an alternative supporting electrolyte was used in enzyme immobilization. Invertase was studied to observe the effect of the supporting electrolyte. Sulfonated poly(arylene ether sulfone) was used as the supporting electrolyte during the electrolysis of pyrrole. The results show that the polymeric supporting electrolyte can be used instead of sodium dodecyl sulfate.
Drug repositioning as an effective therapy for protease-activated receptor 2 inhibition Saqib, Uzma; Savai, Rajkumar; Liu, DongFang; Banerjee, Sreeparna; Baig, Mirza S. (Wiley, 2019-02-01) Proteinase-activated receptor 2 (PAR-2) is a G protein-coupled receptor activated by both trypsin and a specific agonist peptide, SLIGKV-NH2. It has been linked to various pathologies, including pain and inflammation. Several peptide and peptidomimetic agonizts for PAR-2 have been developed exhibiting high potency and efficacy. However, the number of PAR-2 antagonists is smaller. We screened the Food and Drug Administration library of approved compounds to retrieve novel antagonists for repositioning in the...

Citation Formats

S. Kocabıyık, R. Russel, M. Danson, and D. Hough, “The effect of cysteine-43 mutation on thermostability and kinetic properties of citrate synthase from Thermoplasma acidophilum,” BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, pp. 224–228, 1996, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/49070.