Molecular origins of the slow streptavidin-biotin dissociation kinetics

Chilkoti, A; Stayton, P S

doi:10.1021/ja00148a003

Title: Molecular origins of the slow streptavidin-biotin dissociation kinetics

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Abstract

The association of streptavidin and avidin with biotin is among the strongest known noncovalent protein-ligand interactions (K{sub a} nearly equals 2.5 x 10{sup 13} M{sup -1}) and is controlled by an exceptionally slow off-rate. We have used this model system to elucidate the role of aromatic tryptophan side-chain binding contacts in the dissociation reaction coordinate and relatedly to the construction of the activation barrier and to the structure of the transition state. We have also conducted a transition state analysis of the temperature-dependent dissociation kinetics, which along with the independent estimation of the equilibrium biotin-binding free energies and enthalpies has provided thermodynamic profiles defining the enthalpic, entropic, and free energy barriers to dissociation for the mutants relative to wild-type streptavidin. The increased biotin off-rate for W79F, which contacts the valeric acid moiety of biotin, and for W120F, which partially caps the bicyclic ring system, is caused largely by free energy destabilization of the ligand-bound ground state relative to wild-type streptavidin. W79F displays an increased equilibrium binding enthalpy relative to wild-type, and thus streptavidin sacrifices potential binding enthalpy to minimize the entropic costs of biotin immobilization. 26 refs., 5 figs., 4 tabs.

Authors:

Chilkoti, A; Stayton, P S ^[1]

Univ. of Washington, Seattle, WA (United States)

Publication Date:: Wed Nov 01 00:00:00 EST 1995

Sponsoring Org.:: USDOE

OSTI Identifier:: 171510

Resource Type:: Journal Article

Journal Name:: Journal of the American Chemical Society

Additional Journal Information:: Journal Volume: 117; Journal Issue: 43; Other Information: PBD: 1 Nov 1995

Country of Publication:: United States

Language:: English

Subject:: 40 CHEMISTRY; 55 BIOLOGY AND MEDICINE, BASIC STUDIES; 66 PHYSICS; BIOTIN; VITRIFICATION; DISSOCIATION; BINDING ENERGY; CHEMICAL REACTION KINETICS; TEMPERATURE DEPENDENCE; PROTEINS; LIGANDS; GROUND STATES; FREE ENERGY; TRYPTOPHAN

Citation Formats


                    Chilkoti, A, and Stayton, P S. Molecular origins of the slow streptavidin-biotin dissociation kinetics.  United States: N. p., 1995. 
        Web.  doi:10.1021/ja00148a003.

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                    Chilkoti, A, & Stayton, P S. Molecular origins of the slow streptavidin-biotin dissociation kinetics.  United States.  https://doi.org/10.1021/ja00148a003

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                    Chilkoti, A, and Stayton, P S. 1995.  
        "Molecular origins of the slow streptavidin-biotin dissociation kinetics".  United States.  https://doi.org/10.1021/ja00148a003.

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@article{osti_171510,

  title        = {Molecular origins of the slow streptavidin-biotin dissociation kinetics},

  author       = {Chilkoti, A and Stayton, P S},

  abstractNote = {The association of streptavidin and avidin with biotin is among the strongest known noncovalent protein-ligand interactions (K{sub a} nearly equals 2.5 x 10{sup 13} M{sup -1}) and is controlled by an exceptionally slow off-rate. We have used this model system to elucidate the role of aromatic tryptophan side-chain binding contacts in the dissociation reaction coordinate and relatedly to the construction of the activation barrier and to the structure of the transition state. We have also conducted a transition state analysis of the temperature-dependent dissociation kinetics, which along with the independent estimation of the equilibrium biotin-binding free energies and enthalpies has provided thermodynamic profiles defining the enthalpic, entropic, and free energy barriers to dissociation for the mutants relative to wild-type streptavidin. The increased biotin off-rate for W79F, which contacts the valeric acid moiety of biotin, and for W120F, which partially caps the bicyclic ring system, is caused largely by free energy destabilization of the ligand-bound ground state relative to wild-type streptavidin. W79F displays an increased equilibrium binding enthalpy relative to wild-type, and thus streptavidin sacrifices potential binding enthalpy to minimize the entropic costs of biotin immobilization. 26 refs., 5 figs., 4 tabs.},

  doi          = {10.1021/ja00148a003},

  url          = {https://www.osti.gov/biblio/171510},
  journal      = {Journal of the American Chemical Society},
number       = 43,

  volume       = 117,

  place        = {United States},

  year         = {Wed Nov 01 00:00:00 EST 1995},

  month        = {Wed Nov 01 00:00:00 EST 1995}

}

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