The refolding of type II shikimate kinase from<i>Erwinia chrysanthemi</i>after denaturation in urea

Cerasoli, Eleonora; Kelly, Sharon M.; Coggins, John R.; Boam, Deborah J.; Clarke, David T.; Price, Nicholas C.

doi:10.1046/j.1432-1033.2002.ejb.02862.x

Cited by 4 publications

(2 citation statements)

References 45 publications

(57 reference statements)

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“…Chemically-induced unfolding studies usually employ chaotropic agents, such as urea or guanidinium HCl, or hydrophobic additives, such as the detergent sodium dodecyl sulphate (SDS), to interrupt hydrogen bonds and hydrophobic interactions in the interior of proteins. The usefulness of SRCD in folding studies was exemplified in a study of the alpha/beta protein shikimate kinase (Cerasoli et al 2002). This study used a combination of stopped flow studies on a cCD instrument (at the time there were no working SRCD stopped flow setups, see Section 6.1.3) to follow the refolding process, but also used SRCD scans to examine the whole spectrum of the protein in its unfolded state.…”

Section: Chemical Stabilitymentioning

confidence: 99%

Protein characterisation by synchrotron radiation circular dichroism spectroscopy

Wallace

2009

Quart. Rev. Biophys.

102

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Circular dichroism (CD) spectroscopy is a well-established technique for the study of proteins. Synchrotron radiation circular dichroism (SRCD) spectroscopy extends the utility of conventional CD spectroscopy (i.e. using laboratory-based instruments) because the high light flux from a synchrotron enables collection of data to lower wavelengths, detection of spectra with higher signal-to-noise levels and measurements in the presence of strongly absorbing non-chiral components such as salts, buffers, lipids and detergents. This review describes developments in instrumentation, methodologies and bioinformatics that have enabled new applications of the SRCD technique for the study of proteins. It includes examples of the use of SRCD spectroscopy for providing static and dynamic structural information on molecules, including determinations of secondary structures of intact proteins and domains, assessment of protein stability, detection of conformational changes associated with ligand and drug binding, monitoring of environmental effects, examination of the processes of protein folding and membrane insertion, comparisons of mutant and modified proteins, identification of intermolecular interactions and complex formation, determination of the dispositions of proteins in membranes, identification of natively disordered proteins and their binding partners and examination of the carbohydrate components of glycoproteins. It also discusses how SRCD can be used in conjunction with macromolecular crystallography and other biophysical techniques to provide a more complete picture of protein structures and functions, including how proteins interact with other macromolecules and ligands. This review also includes a discussion of potential new applications in structural and functional genomics using SRCD spectroscopy and future instrumentation and bioinformatics developments that will enable such studies. Finally, the appendix describes a number of computational/bioinformatics resources for secondary structure analyses that take advantage of the improved data quality available from SRCD. In summary, this review discusses how SRCD can be used for a wide range of structural and functional studies of proteins.

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Section: Chemical Stabilitymentioning

confidence: 99%

Protein characterisation by synchrotron radiation circular dichroism spectroscopy

Wallace

2009

Quart. Rev. Biophys.

102

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“…While this assay has been widely used for the determination of kinetic parameters of ATP-dependent kinases including SK from different species, − it presents major drawbacks and limitations. First, by monitoring the disappearance of NADH, this assay measures a decrease in signal rather than an increase, resulting in restrictions on the dynamic range of the assay if large amounts of NADH are used.…”

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confidence: 99%

Development of an ESI-LC-MS-Based Assay for Kinetic Evaluation of Mycobacterium tuberculosis Shikimate Kinase Activity and Inhibition

et al. 2015

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A simple and reliable liquid chromatography-mass spectrometry (LC-MS) assay has been developed and validated for the kinetic characterization and evaluation of inhibitors of shikimate kinase from Mycobacterium tuberculosis (MtSK), a potential target for the development of novel antitubercular drugs. This assay is based on the direct determination of the reaction product shikimate-3-phosphate (S3P) using electrospray ionization (ESI) and a quadrupole time-of-flight (Q-TOF) detector. A comparative analysis of the kinetic parameters of MtSK obtained by the LC-MS assay with those obtained by a conventional UV-assay was performed. Kinetic parameters determined by LC-MS were in excellent agreement with those obtained from the UV assay, demonstrating the accuracy, and reliability of this method. The validated assay was successfully applied to the kinetic characterization of a known inhibitor of shikimate kinase; inhibition constants and mode of inhibition were accurately delineated with LC-MS.

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Shikimate kinase

Springer Handbook of Enzymes

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The refolding of type II shikimate kinase fromErwinia chrysanthemiafter denaturation in urea

Cited by 4 publications

References 45 publications

Protein characterisation by synchrotron radiation circular dichroism spectroscopy

Protein characterisation by synchrotron radiation circular dichroism spectroscopy

Development of an ESI-LC-MS-Based Assay for Kinetic Evaluation of Mycobacterium tuberculosis Shikimate Kinase Activity and Inhibition

Shikimate kinase

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