A sequential binding mechanism in a PDZ domain

Research output: Contribution to journalJournal articlepeer-review

Standard

A sequential binding mechanism in a PDZ domain. / Chi, Celestine N; Bach, Anders; Engström, Åke; Wang, Huiqun; Strømgaard, Kristian; Gianni, Stefano; Jemth, Per.

In: Biochemistry, Vol. 48, No. 30, 2009, p. 7089-7097.

Research output: Contribution to journalJournal articlepeer-review

Harvard

Chi, CN, Bach, A, Engström, Å, Wang, H, Strømgaard, K, Gianni, S & Jemth, P 2009, 'A sequential binding mechanism in a PDZ domain', Biochemistry, vol. 48, no. 30, pp. 7089-7097. https://doi.org/10.1021/bi900559k

APA

Chi, C. N., Bach, A., Engström, Å., Wang, H., Strømgaard, K., Gianni, S., & Jemth, P. (2009). A sequential binding mechanism in a PDZ domain. Biochemistry, 48(30), 7089-7097. https://doi.org/10.1021/bi900559k

Vancouver

Chi CN, Bach A, Engström Å, Wang H, Strømgaard K, Gianni S et al. A sequential binding mechanism in a PDZ domain. Biochemistry. 2009;48(30):7089-7097. https://doi.org/10.1021/bi900559k

Author

Chi, Celestine N ; Bach, Anders ; Engström, Åke ; Wang, Huiqun ; Strømgaard, Kristian ; Gianni, Stefano ; Jemth, Per. / A sequential binding mechanism in a PDZ domain. In: Biochemistry. 2009 ; Vol. 48, No. 30. pp. 7089-7097.

Bibtex

@article{d7fff2100fd511df825d000ea68e967b,
title = "A sequential binding mechanism in a PDZ domain",
abstract = "Conformational selection and induced fit are two well-known mechanisms of allosteric protein-ligand interaction. Some proteins, like ubiquitin, have recently been found to exist in multiple conformations at equilibrium, suggesting that the conformational selection may be a general mechanism of interaction, in particular for single-domain proteins. Here, we found that the PDZ2 domain of SAP97 binds its ligand via a sequential (induced fit) mechanism. We performed binding experiments using SAP97 PDZ2 and peptide ligands and observed biphasic kinetics with the stopped-flow technique, indicating that ligand binding involves at least a two-step process. By using an ultrarapid continuous-flow mixer, we then detected a hyperbolic dependence of binding rate constants on peptide concentration, corroborating the two-step binding mechanism. Furthermore, we found a similar dependence of the rate constants on both PDZ and peptide concentration, demonstrating that the PDZ2-peptide interaction involves a precomplex, which then undergoes a conformational change, and thereby follows an induced fit mechanism.",
keywords = "Former Faculty of Pharmaceutical Sciences",
author = "Chi, {Celestine N} and Anders Bach and {\AA}ke Engstr{\"o}m and Huiqun Wang and Kristian Str{\o}mgaard and Stefano Gianni and Per Jemth",
note = "Keywords: Adaptor Proteins, Signal Transducing; Amino Acid Sequence; Binding Sites; DNA-Binding Proteins; Humans; Ligands; Membrane Proteins; Models, Molecular; Molecular Sequence Data; Oncogene Proteins, Viral; PDZ Domains; Peptides; Protein Binding; Protein Conformation; Protein Denaturation; Protein Folding; Thermodynamics",
year = "2009",
doi = "10.1021/bi900559k",
language = "English",
volume = "48",
pages = "7089--7097",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "30",

}

RIS

TY - JOUR

T1 - A sequential binding mechanism in a PDZ domain

AU - Chi, Celestine N

AU - Bach, Anders

AU - Engström, Åke

AU - Wang, Huiqun

AU - Strømgaard, Kristian

AU - Gianni, Stefano

AU - Jemth, Per

N1 - Keywords: Adaptor Proteins, Signal Transducing; Amino Acid Sequence; Binding Sites; DNA-Binding Proteins; Humans; Ligands; Membrane Proteins; Models, Molecular; Molecular Sequence Data; Oncogene Proteins, Viral; PDZ Domains; Peptides; Protein Binding; Protein Conformation; Protein Denaturation; Protein Folding; Thermodynamics

PY - 2009

Y1 - 2009

N2 - Conformational selection and induced fit are two well-known mechanisms of allosteric protein-ligand interaction. Some proteins, like ubiquitin, have recently been found to exist in multiple conformations at equilibrium, suggesting that the conformational selection may be a general mechanism of interaction, in particular for single-domain proteins. Here, we found that the PDZ2 domain of SAP97 binds its ligand via a sequential (induced fit) mechanism. We performed binding experiments using SAP97 PDZ2 and peptide ligands and observed biphasic kinetics with the stopped-flow technique, indicating that ligand binding involves at least a two-step process. By using an ultrarapid continuous-flow mixer, we then detected a hyperbolic dependence of binding rate constants on peptide concentration, corroborating the two-step binding mechanism. Furthermore, we found a similar dependence of the rate constants on both PDZ and peptide concentration, demonstrating that the PDZ2-peptide interaction involves a precomplex, which then undergoes a conformational change, and thereby follows an induced fit mechanism.

AB - Conformational selection and induced fit are two well-known mechanisms of allosteric protein-ligand interaction. Some proteins, like ubiquitin, have recently been found to exist in multiple conformations at equilibrium, suggesting that the conformational selection may be a general mechanism of interaction, in particular for single-domain proteins. Here, we found that the PDZ2 domain of SAP97 binds its ligand via a sequential (induced fit) mechanism. We performed binding experiments using SAP97 PDZ2 and peptide ligands and observed biphasic kinetics with the stopped-flow technique, indicating that ligand binding involves at least a two-step process. By using an ultrarapid continuous-flow mixer, we then detected a hyperbolic dependence of binding rate constants on peptide concentration, corroborating the two-step binding mechanism. Furthermore, we found a similar dependence of the rate constants on both PDZ and peptide concentration, demonstrating that the PDZ2-peptide interaction involves a precomplex, which then undergoes a conformational change, and thereby follows an induced fit mechanism.

KW - Former Faculty of Pharmaceutical Sciences

U2 - 10.1021/bi900559k

DO - 10.1021/bi900559k

M3 - Journal article

C2 - 19496620

VL - 48

SP - 7089

EP - 7097

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 30

ER -

ID: 17365971