PMID- 10514499
OWN - NLM
STAT- MEDLINE
DCOM- 19991119
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 42
DP  - 1999 Oct 15
TI  - Mammal-specific, ERK-dependent, caldesmon phosphorylation in smooth muscle.
      Quantitation using novel anti-phosphopeptide antibodies.
PG  - 30115-21
AB  - Extracellular signal-regulated kinases (ERKs) phosphorylate the high molecular
      mass isoform of the actin-binding protein caldesmon (h-CaD) at two sites
      (Ser(759) and Ser(789)) during smooth muscle stimulation. To investigate the role
      of phosphorylation at these sites, antibodies were generated against
      phosphopeptides analogous to the sequences around Ser(759) and Ser(789).
      Affinity-purified antibodies were phosho- and sequence-specific. The major site
      of phosphorylation in h-CaD in porcine carotid arterial muscle strips was at
      Ser(789); however, the amount of phosphate did not vary appreciably with either
      KCl or phorbol ester stimulation. Phosphorylation at Ser(759) of h-CaD was almost
      undetectable (<0.005 mol of phosphate/mol of protein). Moreover, phosphorylation 
      of the low molecular mass isoform of the protein (l-CaD) at the site analogous to
      Ser(789) was greater in serum-stimulated cultured smooth muscle cells than in
      serum-starved cells. Serum-stimulated l-CaD phosphorylation was attenuated by the
      protein kinase inhibitor PD98059. These data 1) identify Ser(789) of h-CaD as the
      major site of ERK-dependent phosphorylation in carotid arteries; 2) show that the
      level of phosphorylation at Ser(789) is relatively constant following carotid
      arterial muscle stimulation, despite an increase in total protein phosphate
      content; and 3) suggest a functional role for ERK-dependent l-CaD phosphorylation
      in cell division.
FAU - D'Angelo, G
AU  - D'Angelo G
AD  - Boston Biomedical Research Institute, Boston, Massachusetts 02114, USA.
FAU - Graceffa, P
AU  - Graceffa P
FAU - Wang, C A
AU  - Wang CA
FAU - Wrangle, J
AU  - Wrangle J
FAU - Adam, L P
AU  - Adam LP
LA  - eng
GR  - AR30917/AR/NIAMS NIH HHS/United States
GR  - AR41637/AR/NIAMS NIH HHS/United States
GR  - HL56035/HL/NHLBI NIH HHS/United States
GR  - etc.
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Antibodies)
RN  - 0 (Calmodulin-Binding Proteins)
RN  - 0 (Phosphopeptides)
RN  - EC 2.7.11.24 (Mitogen-Activated Protein Kinase 3)
RN  - EC 2.7.11.24 (Mitogen-Activated Protein Kinases)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Antibodies/immunology
MH  - Calmodulin-Binding Proteins/chemistry/*metabolism
MH  - In Vitro Techniques
MH  - Mitogen-Activated Protein Kinase 3
MH  - Mitogen-Activated Protein Kinases/metabolism
MH  - Molecular Sequence Data
MH  - Muscle, Smooth, Vascular/*metabolism
MH  - Phosphopeptides/*immunology
MH  - Phosphorylation
MH  - Sequence Homology, Amino Acid
MH  - Swine
EDAT- 1999/10/09 00:00
MHDA- 1999/10/09 00:01
CRDT- 1999/10/09 00:00
PHST- 1999/10/09 00:00 [pubmed]
PHST- 1999/10/09 00:01 [medline]
PHST- 1999/10/09 00:00 [entrez]
AID - 10.1074/jbc.274.42.30115 [doi]
AID - S0021-9258(19)51872-5 [pii]
PST - ppublish
SO  - J Biol Chem. 1999 Oct 15;274(42):30115-21. doi: 10.1074/jbc.274.42.30115.