ArticleBiophysical journal1999
Crowding effects on EcoRV kinetics and binding.
Article in Biophysical journal, 1999. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 39 papers.
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Who cites it
39 citing papers in PubMed.
- Chemical interactions modulate λProtein science : a publication of the Protein Society · 2023Article
- The Role of Cosolvent-Water Interactions in Effects of the Media on Functionality of Enzymes: A Case Study ofLife (Basel, Switzerland) · 2023Article
- Article
- Preferential Interactions of a Crowder Protein with the Specific Binding Site of a Native Protein Complex.The journal of physical chemistry letters · 2022Article
- Cosolutes Modify Alkaline Phosphatase Catalysis through Osmotic Stress and Crowding Mechanisms.ACS omega · 2021Article
- Macromolecular crowding effects on the kinetics of opposing reactions catalyzed by alcohol dehydrogenase.Biochemistry and biophysics reports · 2021Article
- Article
- Carbohydrate-Based Macromolecular Crowding-Induced Stabilization of Proteins: Towards Understanding the Significance of the Size of the Crowder.Biomolecules · 2019Article
- Effect of Macromolecular Crowding on the FMN-Heme Intraprotein Electron Transfer in Inducible NO Synthase.Biochemistry · 2019Article
- Consequences of Heterogeneous Crowding on an Enzymatic Reaction: A Residence Time Monte Carlo Approach.ACS omega · 2019Article
- Ultrafast dynamics-driven biomolecular recognition where fast activities dictate slow events.Journal of biosciences · 2018Review
- Large cosolutes, small cosolutes, and dihydrofolate reductase activity.Protein science : a publication of the Protein Society · 2017Article
- Characterization of human telomere RNA G-quadruplex structures in vitro and in living cells using 19F NMR spectroscopy.Nucleic acids research · 2017Article
- Macromolecular Crowding Modulates Actomyosin Kinetics.Biophysical journal · 2016Article
- How can macromolecular crowding inhibit biological reactions? The enhanced formation of DNA nanoparticles.Scientific reports · 2016Article
- Interactions of macromolecular crowding agents and cosolutes with small-molecule substrates: effect on horseradish peroxidase activity with two different substrates.The journal of physical chemistry. B · 2014Article
- Crowding activates ClpB and enhances its association with DnaK for efficient protein aggregate reactivation.Biophysical journal · 2014Article
- Improved cell-free RNA and protein synthesis system.PloS one · 2014Article
- Protein-protein interactions in a crowded environment.Biophysical reviews · 2013Review
- Macromolecular crowding as a suppressor of human IAPP fibril formation and cytotoxicity.PloS one · 2013Article
Corrections and comments
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Authors and funding
2 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The cytosol of the cell contains high concentrations of small and large macromolecules, but experimental data are often obtained in dilute solutions that do not reflect in vivo conditions. We have studied the crowding effect that large macromolecules have on EcoRV cleavage by adding high-molecular-weight Ficoll 70 to reaction solutions. Results indicate that Ficoll has surprisingly little effect on overall EcoRV reaction velocity because of offsetting increases in V(max) and K(m), and stronger nonspecific binding. The changes in measured parameters can largely be attributed to the excluded volume effects on reactant activities and the slowing of protein diffusion. Covolume reduction upon binding appears to reinforce nonspecific binding strength, and k(cat) appears to be slowed by stronger nonspecific binding, which slows product release. The data also suggest that effective Ficoll particle volume decreases as its concentration increases above a few weight percent, which may be due to Ficoll interpenetration or compression.
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