ArticleAnalytical biochemistry2014
Crude and purified proteasome activity assays are affected by type of microplate.
Article in Analytical biochemistry, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed.
- Chronic Alcohol Consumption Reprograms Hepatic Metabolism Through Organelle-Specific Acetylation in Mice.Molecular & cellular proteomics : MCP · 2025Article
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- A Ketogenic Diet in Combination with Gemcitabine Mitigates Pancreatic Cancer-Associated Cachexia in Male and Female KPC Mice.International journal of molecular sciences · 2023Article
- Dominant-negative p53-overexpression in skeletal muscle induces cell death and fiber atrophy in rats.Cell death & disease · 2022Article
- NSAIDs-dependent adaption of the mitochondria-proteasome system in immortalized human cardiomyocytes.Scientific reports · 2020Article
- Gender-specific changes in energy metabolism and protein degradation as major pathways affected in livers of mice treated with ibuprofen.Scientific reports · 2020Article
- Diurnal Rhythms Spatially and Temporally Organize Autophagy.Cell reports · 2019Article
- Hepatic Mitochondrial Defects in a Nonalcoholic Fatty Liver Disease Mouse Model Are Associated with Increased Degradation of Oxidative Phosphorylation Subunits.Molecular & cellular proteomics : MCP · 2018Article
- Thermal proteome profiling of breast cancer cells reveals proteasomal activation by CDK4/6 inhibitor palbociclib.The EMBO journal · 2018Article
- Diclofenac induces proteasome and mitochondrial dysfunction in murine cardiomyocytes and hearts.International journal of cardiology · 2016Article
- Delineation of Molecular Pathways Involved in Cardiomyopathies Caused by Troponin T Mutations.Molecular & cellular proteomics : MCP · 2016Article
- Subnormothermic Perfusion in the Isolated Rat Liver Preserves the Antioxidant Glutathione and Enhances the Function of the Ubiquitin Proteasome System.Oxidative medicine and cellular longevity · 2016Article
- The necessity of and strategies for improving confidence in the accuracy of western blots.Expert review of proteomics · 2014Review
- Modelling proteasome and proteasome regulator activities.Biomolecules · 2014Review
- Regulation of ubiquitin-proteasome and autophagy pathways after acute LPS and epoxomicin administration in mice.BMC musculoskeletal disorders · 2014Article
- Genetics of proteasome diseases.Scientifica · 2013Article
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Authors and funding
3 authors.
Funding
Abstract
Measurement of proteasome activity is fast becoming a commonly used assay in many laboratories. The most common method to measure proteasome activity involves measuring the release of fluorescent tags from peptide substrates in black microplates. Comparisons of black plates used for measuring fluorescence with different properties show that the microplate properties significantly affect the measured activities of the proteasome. The microplate that gave the highest reading of trypsin-like activity of the purified 20S proteasome gave the lowest reading of chymotrypsin-like activity of the 20S proteasome. Plates with medium binding surfaces from two different companies showed an approximately 2-fold difference in caspase-like activity for purified 20S proteasomes. Even standard curves generated using free 7-amino-4-methylcoumarin (AMC) were affected by the microplate used. As such, significantly different proteasome activities, as measured in nmol AMC released/mg/min, were obtained for purified 20S proteasomes as well as crude heart and liver samples when using different microplates. The naturally occurring molecule betulinic acid activated the chymotrypsin-like proteasome activity in three different plates but did not affect the proteasome activity in the nonbinding surface microplate. These findings suggest that the type of proteasome activity being measured and sample type are important when selecting a microplate.
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