Regulation of protein kinase C by the cytoskeletal protein calponin

被引:57
作者
Leinweber, B
Parissenti, AM
Gallant, C
Gangopadhyay, SS
Kirwan-Rhude, A
Leavis, PC
Morgan, KG
机构
[1] Boston Biomed Res Inst, Watertown, MA 02472 USA
[2] Northeastern Ontario Reg Canc Ctr, Sudbury, ON P3E 5J1, Canada
[3] Tufts Univ, Grad Sch Biomed Sci, Dept Physiol, Boston, MA 02111 USA
[4] Harvard Univ, Sch Med, Dept Neurol, Boston, MA 02115 USA
[5] Harvard Univ, Sch Med, Dept Med, Boston, MA 02115 USA
关键词
D O I
10.1074/jbc.M008257200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Previous studies from this laboratory have shown that, upon agonist activation, calponin co-immunoprecipitates and co-localizes with protein kinase CE (PKC epsilon) in vascular smooth muscle cells. In the present study we demonstrate that calponin binds directly to the regulatory domain of PKC both in overlay assays and, under native conditions, by sedimentation with lipid vesicles. Calponin was found to bind to the Ca region of both PKC epsilon and PKC alpha with possible involvement of C1B. The C2 region of PKC epsilon binds to the calponin repeats with a requirement for the region between amino acids 160 and 182. We have also found that calponin can directly activate PKC autophosphorylation. By using anti-phospho-antibodies to residue Ser-660 of PKC beta II, we found that calponin, in a lipid-independent manner, increased auto-phosphorylation of PKCa, -epsilon, and -beta II severalfold compared with control conditions. Similarly, calponin was found to increase the amount of P-32-labeled phosphate incorporated into PKC from [gamma-P-32]ATP. We also observed that calponin addition strongly increased the incorporation of radiolabeled phosphate into an exogenous PKC peptide substrate, suggesting an activation of enzyme activity. Thus, these results raise the possibility that calponin may function in smooth muscle to regulate PKC activity by facilitating the phosphorylation of PRC.
引用
收藏
页码:40329 / 40336
页数:8
相关论文
共 44 条
[1]   EFFECT OF CALPONIN ON ACTIN-ACTIVATED MYOSIN ATPASE ACTIVITY [J].
ABE, M ;
TAKAHASHI, K ;
HIWADA, K .
JOURNAL OF BIOCHEMISTRY, 1990, 108 (05) :835-838
[2]   Substrate-dependent activation requirements and kinetic properties of protein kinase C [J].
Andrea, JE ;
Sutherland, C ;
Winter, CK ;
Walsh, MP .
FEBS LETTERS, 1998, 429 (01) :73-77
[3]   Structural comparisons of calponin homology domains: Implications for actin binding [J].
Banuelos, S ;
Saraste, M ;
Carugo, KD .
STRUCTURE, 1998, 6 (11) :1419-1431
[4]   ROLE OF SUBSTRATE IN IMPARTING CALCIUM AND PHOSPHOLIPID REQUIREMENTS TO PROTEIN-KINASE-C ACTIVATION [J].
BAZZI, MD ;
NELSESTUEN, GL .
BIOCHEMISTRY, 1987, 26 (07) :1974-1982
[5]   INTERACTION OF SMOOTH-MUSCLE CALPONIN WITH PHOSPHOLIPIDS [J].
BOGATCHEVA, NV ;
GUSEV, NB .
FEBS LETTERS, 1995, 371 (02) :123-126
[6]   THREONINE-497 IS A CRITICAL SITE FOR PERMISSIVE ACTIVATION OF PROTEIN-KINASE C-ALPHA [J].
CAZAUBON, S ;
BORNANCIN, F ;
PARKER, PJ .
BIOCHEMICAL JOURNAL, 1994, 301 :443-448
[7]   The coatomer protein beta'-COP, a selective binding protein (RACK) for protein kinase C epsilon [J].
Csukai, M ;
Chen, CH ;
DeMatteis, MA ;
MochlyRosen, D .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (46) :29200-29206
[8]   A role for MAP kinase in differentiated smooth muscle contraction evoked by α-adrenoceptor stimulation [J].
Dessy, C ;
Kim, E ;
Sougnez, CL ;
Laporte, R ;
Morgan, KG .
AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY, 1998, 275 (04) :C1081-C1086
[9]   Carboxyl-terminal phosphorylation regulates the function and subcellular localization of protein kinase C βII [J].
Edwards, AS ;
Faux, MC ;
Scott, JD ;
Newton, AC .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (10) :6461-6468
[10]   INTERACTION OF CALPONIN WITH PHOSPHOLIPIDS [J].
FUJII, T ;
YAMANA, K ;
OGOMA, Y ;
KONDO, Y .
JOURNAL OF BIOCHEMISTRY, 1995, 117 (05) :999-1003