Kinetic study of phosphorylation-dependent complex formation between the kinase-inducible domain (KID) of CREB and the KIX domain of CBP on a quartz crystal microbalance

Hisao Matsuno, Hiroyuki Furusawa, Yoshio Okahata

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

We report quantitative analysis of peptide-peptide interactions on a 27 MHz quartz crystal microbalance (QCM) in aqueous solution. The KID (kinase-inducible domain) of transcription factor CREB (cyclic AMP response element binding protein) is known to interact with the KIX domain of coactivator CBP (CREB binding protein), facilitated by phosphorylation at Ser-133 of the KID, The KIX domain peptide (86 aa) was immobilized on the QCM gold electrode surface by means of a poly(ethylene glycol) spacer. Binding of the KID peptide (46 aa) to the KIX peptide was detected by frequency decreases (mass increases) of the QCM. Both maximum binding amount (Δmmax) and association constants (Ka) obtained from the QCM measurements increased as a result of phosphorylation of Ser-133 of the KID peptide. The Ka values for KIX peptide to the phosphorylated (pKID) and unphosphorylated KID peptides were (93±2)×103 and (5± 1) × 103M-1, respectively. This difference was explained by the dissociation rate constant (k-1) of the pKID being 20 times smaller than that of the KID, while association rate constants (κ1) were independent of phosphorylation.

Original languageEnglish
Pages (from-to)6172-6178
Number of pages7
JournalChemistry - A European Journal
Volume10
Issue number23
DOIs
Publication statusPublished - Nov 19 2004

Fingerprint

CREB-Binding Protein
Cyclic AMP Response Element-Binding Protein
Phosphorylation
Quartz crystal microbalances
Peptides
Phosphotransferases
Kinetics
Rate constants
Association reactions
Transcription factors
Carrier Proteins
Cyclic AMP
Gold
Polyethylene glycols
Transcription Factors
Electrodes

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Chemistry(all)
  • Organic Chemistry

Cite this

@article{5910bb353c6b4f13a7edc611d6762add,
title = "Kinetic study of phosphorylation-dependent complex formation between the kinase-inducible domain (KID) of CREB and the KIX domain of CBP on a quartz crystal microbalance",
abstract = "We report quantitative analysis of peptide-peptide interactions on a 27 MHz quartz crystal microbalance (QCM) in aqueous solution. The KID (kinase-inducible domain) of transcription factor CREB (cyclic AMP response element binding protein) is known to interact with the KIX domain of coactivator CBP (CREB binding protein), facilitated by phosphorylation at Ser-133 of the KID, The KIX domain peptide (86 aa) was immobilized on the QCM gold electrode surface by means of a poly(ethylene glycol) spacer. Binding of the KID peptide (46 aa) to the KIX peptide was detected by frequency decreases (mass increases) of the QCM. Both maximum binding amount (Δmmax) and association constants (Ka) obtained from the QCM measurements increased as a result of phosphorylation of Ser-133 of the KID peptide. The Ka values for KIX peptide to the phosphorylated (pKID) and unphosphorylated KID peptides were (93±2)×103 and (5± 1) × 103M-1, respectively. This difference was explained by the dissociation rate constant (k-1) of the pKID being 20 times smaller than that of the KID, while association rate constants (κ1) were independent of phosphorylation.",
author = "Hisao Matsuno and Hiroyuki Furusawa and Yoshio Okahata",
year = "2004",
month = "11",
day = "19",
doi = "10.1002/chem.200400030",
language = "English",
volume = "10",
pages = "6172--6178",
journal = "Chemistry - A European Journal",
issn = "0947-6539",
publisher = "Wiley-VCH Verlag",
number = "23",

}

TY - JOUR

T1 - Kinetic study of phosphorylation-dependent complex formation between the kinase-inducible domain (KID) of CREB and the KIX domain of CBP on a quartz crystal microbalance

AU - Matsuno, Hisao

AU - Furusawa, Hiroyuki

AU - Okahata, Yoshio

PY - 2004/11/19

Y1 - 2004/11/19

N2 - We report quantitative analysis of peptide-peptide interactions on a 27 MHz quartz crystal microbalance (QCM) in aqueous solution. The KID (kinase-inducible domain) of transcription factor CREB (cyclic AMP response element binding protein) is known to interact with the KIX domain of coactivator CBP (CREB binding protein), facilitated by phosphorylation at Ser-133 of the KID, The KIX domain peptide (86 aa) was immobilized on the QCM gold electrode surface by means of a poly(ethylene glycol) spacer. Binding of the KID peptide (46 aa) to the KIX peptide was detected by frequency decreases (mass increases) of the QCM. Both maximum binding amount (Δmmax) and association constants (Ka) obtained from the QCM measurements increased as a result of phosphorylation of Ser-133 of the KID peptide. The Ka values for KIX peptide to the phosphorylated (pKID) and unphosphorylated KID peptides were (93±2)×103 and (5± 1) × 103M-1, respectively. This difference was explained by the dissociation rate constant (k-1) of the pKID being 20 times smaller than that of the KID, while association rate constants (κ1) were independent of phosphorylation.

AB - We report quantitative analysis of peptide-peptide interactions on a 27 MHz quartz crystal microbalance (QCM) in aqueous solution. The KID (kinase-inducible domain) of transcription factor CREB (cyclic AMP response element binding protein) is known to interact with the KIX domain of coactivator CBP (CREB binding protein), facilitated by phosphorylation at Ser-133 of the KID, The KIX domain peptide (86 aa) was immobilized on the QCM gold electrode surface by means of a poly(ethylene glycol) spacer. Binding of the KID peptide (46 aa) to the KIX peptide was detected by frequency decreases (mass increases) of the QCM. Both maximum binding amount (Δmmax) and association constants (Ka) obtained from the QCM measurements increased as a result of phosphorylation of Ser-133 of the KID peptide. The Ka values for KIX peptide to the phosphorylated (pKID) and unphosphorylated KID peptides were (93±2)×103 and (5± 1) × 103M-1, respectively. This difference was explained by the dissociation rate constant (k-1) of the pKID being 20 times smaller than that of the KID, while association rate constants (κ1) were independent of phosphorylation.

UR - http://www.scopus.com/inward/record.url?scp=9644288158&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=9644288158&partnerID=8YFLogxK

U2 - 10.1002/chem.200400030

DO - 10.1002/chem.200400030

M3 - Article

VL - 10

SP - 6172

EP - 6178

JO - Chemistry - A European Journal

JF - Chemistry - A European Journal

SN - 0947-6539

IS - 23

ER -