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 language | English |
|---|---|
| Pages (from-to) | 6172-6178 |
| Number of pages | 7 |
| Journal | Chemistry - A European Journal |
| Volume | 10 |
| Issue number | 23 |
| DOIs | |
| Publication status | Published - Nov 19 2004 |
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All Science Journal Classification (ASJC) codes
- Catalysis
- Chemistry(all)
- Organic Chemistry
Cite this
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. / Matsuno, Hisao; Furusawa, Hiroyuki; Okahata, Yoshio.
In: Chemistry - A European Journal, Vol. 10, No. 23, 19.11.2004, p. 6172-6178.Research output: Contribution to journal › Article
}
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.
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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 -