ArticleAnalytical biochemistry2010
Use of thermal melt curves to assess the quality of enzyme preparations.
Article in Analytical biochemistry, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed.
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- Functional and in silico analysis of ATP8A2 and other P4-ATPase variants associated with human genetic diseases.Disease models & mechanisms · 2024Article
- Viral Receptor-Binding Protein Evolves New Function through Mutations That Cause Trimer Instability and Functional Heterogeneity.Molecular biology and evolution · 2024Article
- Fluorescence-Based Protein Stability Monitoring-A Review.International journal of molecular sciences · 2024Review
- Purification and characterization of cysteine protease of Sarcocystis fusiformis from infected Egyptian water buffaloes.Scientific reports · 2023Article
- Catalytic Properties of Caseinolytic Protease Subunit ofMolecules (Basel, Switzerland) · 2022Article
- Methods for the Discovery and Identification of Small Molecules Targeting Oxidative Stress-Related Protein-Protein Interactions: An Update.Antioxidants (Basel, Switzerland) · 2022Review
- Standard operating procedure for fluorescent thermal shift assay (FTSA) for determination of protein-ligand binding and protein stability.European biophysics journal : EBJ · 2021Article
- Pocket detection and interaction-weighted ligand-similarity search yields novel high-affinity binders for Myocilin-OLF, a protein implicated in glaucoma.Bioorganic & medicinal chemistry letters · 2017Article
- Identification and Validation of Small-Gatekeeper Kinases as Drug Targets in Giardia lamblia.PLoS neglected tropical diseases · 2016Article
- Novel small molecule binders of human N-glycanase 1, a key player in the endoplasmic reticulum associated degradation pathway.Bioorganic & medicinal chemistry · 2016Article
- Biochemical Screening of Five Protein Kinases from Plasmodium falciparum against 14,000 Cell-Active Compounds.PloS one · 2016Article
- PoLi: A Virtual Screening Pipeline Based on Template Pocket and Ligand Similarity.Journal of chemical information and modeling · 2015Article
- Article
- Analysis of protein stability and ligand interactions by thermal shift assay.Current protocols in protein science · 2015Article
- Ligand induced stabilization of the melting temperature of the HSV-1 single-strand DNA binding protein using the thermal shift assay.Biochemical and biophysical research communications · 2014Article
- Experimental validation of FINDSITE(comb) virtual ligand screening results for eight proteins yields novel nanomolar and micromolar binders.Journal of cheminformatics · 2014Article
- Recent developments in the use of differential scanning fluorometry in protein and small molecule discovery and characterization.Expert opinion on drug discovery · 2013Review
- Structure of hepatitis C virus polymerase in complex with primer-template RNA.Journal of virology · 2012Article
- Crystal structure of GAP50, the anchor of the invasion machinery in the inner membrane complex of Plasmodium falciparum.Journal of structural biology · 2012Article
Corrections and comments
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Authors and funding
12 authors.
Funding
Abstract
This study sought to determine whether the quality of enzyme preparations can be determined from their melting curves, which may easily be obtained using a fluorescent probe and a standard reverse transcription-polymerase chain reaction (RT-PCR) machine. Thermal melt data on 31 recombinant enzymes from Plasmodium parasites were acquired by incrementally heating them to 90 degrees C and measuring unfolding with a fluorescent dye. Activity assays specific to each enzyme were also performed. Four of the enzymes were denatured to varying degrees with heat and sodium dodecyl sulfate (SDS) prior to the thermal melt and activity assays. In general, melting curve quality was correlated with enzyme activity; enzymes with high-quality curves were found almost uniformly to be active, whereas those with lower quality curves were more varied in their catalytic performance. Inspection of melting curves of bovine xanthine oxidase and Entamoeba histolytica cysteine protease 1 allowed active stocks to be distinguished from inactive stocks, implying that a relationship between melting curve quality and activity persists over a wide range of experimental conditions and species. Our data suggest that melting curves can help to distinguish properly folded proteins from denatured ones and, therefore, may be useful in selecting stocks for further study and in optimizing purification procedures for specific proteins.
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