Abstract
The substrate recognition determinants of Ca2+ -calmodulin-dependent protein kinase (CaMK) IV and CaMKIIα were investigated using peptide substrates modeled on the amino acid sequence encompassing Set-9 of synapsin I. For both kinases, hydrophobic residues (Leu or Phe) at the -5 position, are well tolerated, whereas non-hydrophobic residues (Arg, Ala, or Asp) decrease V(max)/K(m) by 55- to >4000-fold. At the -3 position, substitution of Ala for Arg leads to decreases of 99-and 343- fold in V(max)/K(m) for CaMKIV and CaMKIIα, respectively. For both kinases, the nature of the residues occupying the -4, -1, and +4 positions exerts relatively little influence on phosphorylation kinetics. CaMKIV and CaMKIIα respond differently to substitutions at the -2 and +1 positions. Substitution of Arg at the -2 position with non-basic residues (Gln or Ala) leads to 6-fold decreases in V(max)/K(m) for CaMKIV, but 17-28-fold increases for CaMKIIα. Additionally, peptides containing Leu, Asp, or Ala at the +1 position are phosphorylated with similar efficiencies by CaMKIV, whereas the Leu- substituted peptide is preferred by CaMKIIα (by a factor of 5.8-9.7-fold). Thus, CaMKIV and CaMKHα preferentially phosphorylate substrates with the motifs: Hyd-X-Arg-X-X-Ser*/Thr*, and Hyd-X-Arg-NB-X-Ser*/Thr*-Hyd, respectively, where Hyd represents a hydrophobic, X any, and NB a non-basic amino acid residue. The different specificities of the two kinases may contribute to their targeting to distinct physiological substrates during Ca2+ -dependent cellular events.
| Original language | English |
|---|---|
| Pages (from-to) | 3166-3172 |
| Number of pages | 7 |
| Journal | Journal of Biological Chemistry |
| Volume | 273 |
| Issue number | 6 |
| DOIs | |
| Publication status | Published - 1998 Feb 6 |
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All Science Journal Classification (ASJC) codes
- Biochemistry
- Molecular Biology
- Cell Biology
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Definition of optimal substrate recognition motifs of Ca2+- calmodulin-dependent protein kinases IV and II reveals shared and distinctive features. / White, Ronald R.; Kwon, Young Guen; Taing, Meng; Lawrence, David S.; Edelman, Arthur M.
In: Journal of Biological Chemistry, Vol. 273, No. 6, 06.02.1998, p. 3166-3172.Research output: Contribution to journal › Article
TY - JOUR
T1 - Definition of optimal substrate recognition motifs of Ca2+- calmodulin-dependent protein kinases IV and II reveals shared and distinctive features
AU - White, Ronald R.
AU - Kwon, Young Guen
AU - Taing, Meng
AU - Lawrence, David S.
AU - Edelman, Arthur M.
PY - 1998/2/6
Y1 - 1998/2/6
N2 - The substrate recognition determinants of Ca2+ -calmodulin-dependent protein kinase (CaMK) IV and CaMKIIα were investigated using peptide substrates modeled on the amino acid sequence encompassing Set-9 of synapsin I. For both kinases, hydrophobic residues (Leu or Phe) at the -5 position, are well tolerated, whereas non-hydrophobic residues (Arg, Ala, or Asp) decrease V(max)/K(m) by 55- to >4000-fold. At the -3 position, substitution of Ala for Arg leads to decreases of 99-and 343- fold in V(max)/K(m) for CaMKIV and CaMKIIα, respectively. For both kinases, the nature of the residues occupying the -4, -1, and +4 positions exerts relatively little influence on phosphorylation kinetics. CaMKIV and CaMKIIα respond differently to substitutions at the -2 and +1 positions. Substitution of Arg at the -2 position with non-basic residues (Gln or Ala) leads to 6-fold decreases in V(max)/K(m) for CaMKIV, but 17-28-fold increases for CaMKIIα. Additionally, peptides containing Leu, Asp, or Ala at the +1 position are phosphorylated with similar efficiencies by CaMKIV, whereas the Leu- substituted peptide is preferred by CaMKIIα (by a factor of 5.8-9.7-fold). Thus, CaMKIV and CaMKHα preferentially phosphorylate substrates with the motifs: Hyd-X-Arg-X-X-Ser*/Thr*, and Hyd-X-Arg-NB-X-Ser*/Thr*-Hyd, respectively, where Hyd represents a hydrophobic, X any, and NB a non-basic amino acid residue. The different specificities of the two kinases may contribute to their targeting to distinct physiological substrates during Ca2+ -dependent cellular events.
AB - The substrate recognition determinants of Ca2+ -calmodulin-dependent protein kinase (CaMK) IV and CaMKIIα were investigated using peptide substrates modeled on the amino acid sequence encompassing Set-9 of synapsin I. For both kinases, hydrophobic residues (Leu or Phe) at the -5 position, are well tolerated, whereas non-hydrophobic residues (Arg, Ala, or Asp) decrease V(max)/K(m) by 55- to >4000-fold. At the -3 position, substitution of Ala for Arg leads to decreases of 99-and 343- fold in V(max)/K(m) for CaMKIV and CaMKIIα, respectively. For both kinases, the nature of the residues occupying the -4, -1, and +4 positions exerts relatively little influence on phosphorylation kinetics. CaMKIV and CaMKIIα respond differently to substitutions at the -2 and +1 positions. Substitution of Arg at the -2 position with non-basic residues (Gln or Ala) leads to 6-fold decreases in V(max)/K(m) for CaMKIV, but 17-28-fold increases for CaMKIIα. Additionally, peptides containing Leu, Asp, or Ala at the +1 position are phosphorylated with similar efficiencies by CaMKIV, whereas the Leu- substituted peptide is preferred by CaMKIIα (by a factor of 5.8-9.7-fold). Thus, CaMKIV and CaMKHα preferentially phosphorylate substrates with the motifs: Hyd-X-Arg-X-X-Ser*/Thr*, and Hyd-X-Arg-NB-X-Ser*/Thr*-Hyd, respectively, where Hyd represents a hydrophobic, X any, and NB a non-basic amino acid residue. The different specificities of the two kinases may contribute to their targeting to distinct physiological substrates during Ca2+ -dependent cellular events.
UR - http://www.scopus.com/inward/record.url?scp=0032488899&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0032488899&partnerID=8YFLogxK
U2 - 10.1074/jbc.273.6.3166
DO - 10.1074/jbc.273.6.3166
M3 - Article
C2 - 9452427
AN - SCOPUS:0032488899
VL - 273
SP - 3166
EP - 3172
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 6
ER -