Proteomic analysis of IgE-mediated secretion by LAD2 mast cells.
The LAD2 cell line is a relatively recent addition to the range of mast cell analogues and is of particular importance as it is the only human analogue which can be stimulated to degranulate in an IgE-dependent manner. Mast cells are tissue-based effector cells which have historically been shown to play an important role in the adaptive immune response, though there is now gathering evidence of their significance as a component of the innate immune system. These functions can be attributed to the ability of mast cells to regulate secretion of a wide variety of potent biologically active mediators through immediate and delayed responses. This well-orchestrated secretory mechanism of the mast cell makes it an ideal model in which to study this event. In this investigation, two-dimensional electrophoresis was employed as part of the proteomic characterization of the LAD2 human mast cell line, focusing in particular on a global analysis of membrane protein relocation after an IgE-mediated stimulatory event. This investigation has identified six membrane-associated protein spots which became phosphorylated upon IgE-mediated activation, 31 protein spots which displayed consistent recruitment to the membrane fraction, and three which were consistently lost from the soluble fraction. The scenario which emerges reveals a series of substantial changes which affect every compartment of the cell, providing evidence for a coordinated response to a secretory stimulus.
Gage
Matthew C
MC
Institute of Membrane and Systems Biology, University of Leeds, United Kingdom.
Keen
Jeffrey N
JN
Buxton
Anthony T
AT
Bedi
Maninder K
MK
Findlay
John B C
JB
D015550:Cell Degranulation; D002460:Cell Line; D018118:Chloride Channels / Q000378:metabolism; D002853:Chromatography, Liquid; D003598:Cytoskeletal Proteins / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D005089:Exocytosis; D006801:Humans; D007073:Immunoglobulin E / Q000378:metabolism*; D008407:Mast Cells / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism; D010766:Phosphorylation; D040901:Proteomics / Q000379:methods*; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction; D012995:Solubility; D053719:Tandem Mass Spectrometry
D018118:Chloride Channels; D003598:Cytoskeletal Proteins; D008565:Membrane Proteins; D017382:Reactive Oxygen Species; D007073:Immunoglobulin E