ArticleBiophysical journal1994
Resolution of multiphasic reactions by the combination of fluorescence total-intensity and anisotropy stopped-flow kinetic experiments.
Article in Biophysical journal, 1994. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed.
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- Hsp104 and Potentiated Variants Can Operate as Distinct Nonprocessive Translocases.Biophysical journal · 2019Article
- Detailed characterization of the solution kinetics and thermodynamics of biotin, biocytin and HABA binding to avidin and streptavidin.PloS one · 2019Article
- The fluorescence intensities ratio is not a reliable parameter for evaluation of protein unfolding transitions.Protein science : a publication of the Protein Society · 2017Article
- Modeling the Actin.myosin ATPase Cross-Bridge Cycle for Skeletal and Cardiac Muscle Myosin Isoforms.Biophysical journal · 2017Article
- Article
- Stability of the Plasmodium falciparum AMA1-RON2 Complex Is Governed by the Domain II (DII) Loop.PloS one · 2016Article
- Global multi-method analysis of affinities and cooperativity in complex systems of macromolecular interactions.Analytical chemistry · 2012Article
- Kinetic analysis of autotaxin reveals substrate-specific catalytic pathways and a mechanism for lysophosphatidic acid distribution.The Journal of biological chemistry · 2011Article
- Pathway of ATP utilization and duplex rRNA unwinding by the DEAD-box helicase, DbpA.Proceedings of the National Academy of Sciences of the United States of America · 2010Article
- Dynamics of the ssDNA recognition by the RepA hexameric helicase of plasmid RSF1010: analyses using fluorescence stopped-flow intensity and anisotropy methods.Journal of molecular biology · 2009Article
- Substrate dependence of conformational changes in the RNA-binding domain of tristetraprolin assessed by fluorescence spectroscopy of tryptophan mutants.Biochemistry · 2006Article
- Asymmetric effect of domain interactions on the kinetics of folding in yeast phosphoglycerate kinase.Protein science : a publication of the Protein Society · 2005Article
- Proline can have opposite effects on fast and slow protein folding phases.Biophysical journal · 2003Article
- Application of fluorescence resonance energy transfer to the GroEL-GroES chaperonin reaction.Methods (San Diego, Calif.) · 2001Review
- High-sensitivity fluorescence anisotropy detection of protein-folding events: application to alpha-lactalbumin.Biophysical journal · 2001Article
- ClpA mediates directional translocation of substrate proteins into the ClpP protease.Proceedings of the National Academy of Sciences of the United States of America · 2001Article
- A fluorescence-based assay for 3' --> 5' exoribonucleases: potential applications to the study of mRNA decay.RNA (New York, N.Y.) · 2000Article
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3 authors.
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Abstract
Multiphasic kinetics are often observed in stopped-flow investigations. To characterize further these kinetic phases, we have developed a methodology whereby fluorescence total intensity and anisotropy stopped-flow data can be combined in a single analysis. Fluorescence total intensity and anisotropy are highly interrelated and contain two very complementary forms of information. Total-intensity changes are useful in determining changes in populations with differing quantum yields, whereas anisotropy changes contain additional contributions caused by the rotational dynamics of the species. For cases in which the fluorescence quantum yield increases, the observed rate of anisotropy change will be more rapid than the total-intensity change, whereas in cases in which the total intensity decreases, the observed change in anisotropy will lag behind. In all cases, with quantum yield changes the stopped-flow anisotropy signals cannot be fit with models consisting of exponentials. Case studies examining these effects are described for the protein folding/refolding transitions of Staphylococcal nuclease and phosphoglycerate kinase. A multiphasic DNA exonuclease reaction using bacteriophage T4 DNA polymerase is also examined. In all of these cases, combined analysis of both data types revealed insights into reaction mechanism, which could not be obtained by either data type in isolation. Quantum yields and steady-state anisotropies associated with transiently populated intermediate species can be resolved. The data analysis methodologies described allow characterization of multiphasic reactions in terms of internally consistent kinetic rates, quantum yields, and steady-state anisotropies.
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