Parallel single-cell monitoring of receptor-triggered membrane translocation of a calcium-sensing protein module.
Time courses of translocation of fluorescently conjugated proteins to the plasma membrane were simultaneously measured in thousands of individual rat basophilic leukemia cells. We found that the C2 domain---a calcium-sensing, lipid-binding protein module that is an essential regulator of protein kinase C and numerous other proteins---targeted proteins to the plasma membrane transiently if calcium was released from internal stores, and persistently in response to entry of extracellular calcium across the plasma membrane. The C2 domain translocation time courses of stimulated cells clustered into only two primary modes. Hence, the reversible recruitment of families of signaling proteins from one cellular compartment to another is a rapid bifurcation mechanism for inducing discrete states of cellular signaling networks.
Teruel
Mary N
MN
Department of Molecular Pharmacology, Stanford University Medical School, 269 Campus Drive, Stanford, CA 94305, USA.
Meyer
Tobias
T
D000818:Animals; D001426:Bacterial Proteins; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling; D002462:Cell Membrane / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D005453:Fluorescence; D005456:Fluorescent Dyes; D007527:Isoenzymes / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D008164:Luminescent Proteins; D010972:Platelet Activating Factor / Q000494:pharmacology; D011485:Protein Binding; D011493:Protein Kinase C / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D051381:Rats; D011956:Receptors, Cell Surface / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012984:Software; D019284:Thapsigargin / Q000494:pharmacology; D014162:Transfection; D014407:Tumor Cells, Cultured
D001426:Bacterial Proteins; D005456:Fluorescent Dyes; D007527:Isoenzymes; D008164:Luminescent Proteins; D010972:Platelet Activating Factor; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; C054562:yellow fluorescent protein, Bacteria; D019284:Thapsigargin; C087857:protein kinase C gamma; D011493:Protein Kinase C; D002118:Calcium