Abstract

Our previous studies indicated that IgE-mediated stimulation of rat baso- philic leukemia (RBL-2H3) cells resulted in the secretion of histamine and the phosphorylation of the heavy (200 kDa) and light (20 kDa) chains of myosin. In the case of the 20-kDa myosin light chain, all of the phosphate was confined to a single site, which could be identified as the serine residue phosphorylated by MLCK in both turkey gizzard smooth muscle and human platelet myosin. Following RBL-2H3 cell stimulation there was de novo phosphorylation of a serine site that had previously been shown to be phosphorylated by protein kinase C in turkey gizzard and human platelet myosin. A similar scenario was observed for the myosin heavy chain. Unstimulated cells contained myosin heavy chains that yielded a number of phosphorylate tryptic peptides. However, following antigen stimulation, a single new tryptic phosphopeptide appeared. This peptide appeared to be one whose phosphorylation was catalyzed by protein kinase C. The stoichiometry of phosphorylation of these sites was substantial (> 0.5 moles of phosphate/myosin, light and heavy chain) and was closely correlated with the extent of exocytosis.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Intramural Research (Z01)
Project #
1Z01HL000993-04
Application #
3878916
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
4
Fiscal Year
1990
Total Cost
Indirect Cost

  Institution

Name
National Heart, Lung, and Blood Institute
Department
Type
DUNS #
City
State
Country
United States
Zip Code

  Publications

Park, Seung-Kiel; Qiao, Huihong; Beaven, Michael A (2009) Src-like adaptor protein (SLAP) is upregulated in antigen-stimulated mast cells and acts as a negative regulator. Mol Immunol 46:2133-9
Ma, Hong-Tao; Peng, Ze; Hiragun, Takaaki et al. (2008) Canonical transient receptor potential 5 channel in conjunction with Orai1 and STIM1 allows Sr2+ entry, optimal influx of Ca2+, and degranulation in a rat mast cell line. J Immunol 180:2233-9
Beaven, Michael A (2007) Division of labor: specialization of sphingosine kinases in mast cells. Immunity 26:271-3
Mitra, Poulami; Oskeritzian, Carole A; Payne, Shawn G et al. (2006) Role of ABCC1 in export of sphingosine-1-phosphate from mast cells. Proc Natl Acad Sci U S A 103:16394-9
Lee, Jun Ho; Kim, Young Mi; Kim, Nam Wook et al. (2006) Phospholipase D2 acts as an essential adaptor protein in the activation of Syk in antigen-stimulated mast cells. Blood 108:956-64
Iwaki, Shoko; Tkaczyk, Christine; Satterthwaite, Anne B et al. (2005) Btk plays a crucial role in the amplification of Fc epsilonRI-mediated mast cell activation by kit. J Biol Chem 280:40261-70
Peng, Ze; Beaven, Michael A (2005) An essential role for phospholipase D in the activation of protein kinase C and degranulation in mast cells. J Immunol 174:5201-8
Andrade, Marcus V M; Hiragun, Takaaki; Beaven, Michael A (2004) Dexamethasone suppresses antigen-induced activation of phosphatidylinositol 3-kinase and downstream responses in mast cells. J Immunol 172:7254-62
Choi, Wahn Soo; Hiragun, Takaaki; Lee, Jun Ho et al. (2004) Activation of RBL-2H3 mast cells is dependent on tyrosine phosphorylation of phospholipase D2 by Fyn and Fgr. Mol Cell Biol 24:6980-92
Tkaczyk, Christine; Beaven, Michael A; Brachman, Saskia M et al. (2003) The phospholipase C gamma 1-dependent pathway of Fc epsilon RI-mediated mast cell activation is regulated independently of phosphatidylinositol 3-kinase. J Biol Chem 278:48474-84

Showing the most recent 10 out of 15 publications