ArticleProtein science : a publication of the Protein Society2007
Analysis of ligand binding to a ribose biosensor using site-directed mutagenesis and fluorescence spectroscopy.
Article in Protein science : a publication of the Protein Society, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 35 citations in OpenAlex.
- Ribose-Binding Protein Mutants With Improved Interaction Towards the Non-natural Ligand 1,3-Cyclohexanediol.Frontiers in bioengineering and biotechnology · 2021Article
- Ligand-induced structural changes analysis of ribose-binding protein as studied by molecular dynamics simulations.Technology and health care : official journal of the European Society for Engineering and Medicine · 2021Article
- Computational redesign of the Escherichia coli ribose-binding protein ligand binding pocket for 1,3-cyclohexanediol and cyclohexanol.Scientific reports · 2019Article
- A Single Protein Disruption Site Results in Efficient Reassembly by Multiple Engineering Methods.Biophysical journal · 2019Article
- Complete alanine scanning of the Escherichia coli RbsB ribose binding protein reveals residues important for chemoreceptor signaling and periplasmic abundance.Scientific reports · 2017Article
- Small Molecule-Induced Domain Swapping as a Mechanism for Controlling Protein Function and Assembly.Scientific reports · 2017Article
- Molecular Insights into the Potential Toxicological Interaction of 2-Mercaptothiazoline with the Antioxidant Enzyme-Catalase.International journal of molecular sciences · 2016Article
- Strep-tag II Mutant Maltose-binding Protein for Reagentless Fluorescence Sensing.Tropical life sciences research · 2016Article
- Engineered Domain Swapping as an On/Off Switch for Protein Function.Chemistry & biology · 2015Article
- Analysis of conformational motions and residue fluctuations for Escherichia coli ribose-binding protein revealed with elastic network models.International journal of molecular sciences · 2013Article
- Stepwise conversion of a binding protein to a fluorescent switch: application to Thermoanaerobacter tengcongensis ribose binding protein.Biochemistry · 2013Article
- Protein conformational switches: from nature to design.Chemistry (Weinheim an der Bergstrasse, Germany) · 2012Review
- Computational design of ligand binding is not a solved problem.Proceedings of the National Academy of Sciences of the United States of America · 2009Article
- Computational design of Candida boidinii xylose reductase for altered cofactor specificity.Protein science : a publication of the Protein Society · 2009Article
- Beyond molecular beacons: optical sensors based on the binding-induced folding of proteins and polypeptides.Chemistry (Weinheim an der Bergstrasse, Germany) · 2009Article
- A Ca2+-sensing molecular switch based on alternate frame protein folding.ACS chemical biology · 2008Article
- Ligand-induced conformational changes in a thermophilic ribose-binding protein.BMC structural biology · 2008Article
Corrections and comments
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Computational design of proteins with altered ligand specificity is an emerging method for the creation of new biosensing systems. In this work, we investigated the outcome of site-directed mutagenesis on the Escherichia coli ribose binding protein (RBP), which is frequently used as a design scaffold for computational searches. A ribose biosensor was first constructed whereby an environmentally sensitive fluorescent probe was covalently attached to RBP at position S265C. This protein conjugate displayed a 54% decrease in emission intensity upon the addition of saturating ribose concentrations and exhibited an apparent dissociation constant (K(d) ) of 3.4 microM. Site-directed mutants within the RBP binding pocket were created and examined for ribose binding ability and overall structural stability. Because as many as 12 mutations are needed to alter ligand specificity in RBP, we measured the effect of single and multiple alanine mutations on stability and signal transduction potential of the ribose biosensor. Single alanine mutations had significant impact on both stability and signaling. Mutations of N190A and F214A each produced melting temperatures >8 degrees C below those observed for the wild-type protein. Residue Q235, located in the hinge region of RBP, appeared to be a hot spot for global protein stability as well. Additional single alanine mutations demonstrated as much as 200-fold increase in apparent K(d) but retained overall protein stability. The data collected from this study may be incorporated into design algorithms to help create more stable biosensors and optimize signal transduction properties for a variety of important analytes.
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