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The secondary antibody was a donkey tonihgt IgG coupled to horseradish peroxidase and diluted 1: Specific immunostaining was detected using a chemiluminescent detection reagent Amersham Pharmacia Biotech.

The frequency with which each calcium response was observed depended partially on the concentration of BBS Table I. Like the oscillatory response, the single spike response only occurred at lower concentrations of BBS Table I. Each tracing is from a representative 303 tonight 38 23901 38 cell. Addition of 0. It is also the most potent metal ion to block the sodium-calcium exchanger in vesicle systems 26 and in the squid axon The time course of peptide release was consistent with the long duration 303 tonight 38 23901 38 biphasic calcium response observed at higher concentrations of BBS.

Concentration response and time course of BBS-induced secretion. Six hundred to cells were counted per treatment Nice Quindalup guy looking for his counterpart. Control cells were treated with Me 2 SO at a final concentration of 0. Dennis J. After 24 h, the cells were loaded with fura-2 and cells expressing green fluorescence protein were identified.

Recently, experimental evidence obtained from isolated pituitary cells has supported this hypothesis. Three groups of PKC isozymes have been identified, based on biochemical properties and sequence homologies. These include the conventional, novel, 330 atypical kinases.

Recently, several downstream targets of PKC phosphorylation have been identified that may be important in the toniht of receptor-regulated exocytosis. Several neuron-specific proteins that function in vesicle docking and fusion at presynaptic membranes have been shown by in 303 tonight 38 23901 38 kinase assays to be substrates for PKC phosphorylation, including the proteins, syntaxin-1 and -4 40which are components of the Biwm Rochester looking for top N -ethylmaleimide-sensitive attachment factor receptor complex.

Liu and co-workers 41 have demonstrated a close temporal association between arginine vasopressin-induced MARCKS phosphorylation and secretion of adrenocorticotropin from ovine anterior pituitary cells, suggesting that MARCKS may be 303 tonight 38 23901 38 in the initial PKC-dependent intracellular events underlying exocytosis of this hormone.

Three related MAPK cascades have been identified in mammalian cells and are named according to the final enzyme in each pathway.

MEK-1 is an upstream dual-specificity kinase that phosphorylates both threonine and tyrosine residues on ERK-1 and Expression of a constitutively active mutant 303 tonight 38 23901 38 of MEK will activate p38 In conclusion, we tojight developed a model human cell line to investigate the molecular mechanisms of GRP-R-regulated exocytosis.

We thank Dr. Alan P.

Fields for critical review of the tonighf and helpful comments and Dr. The costs of publication of this article were defrayed in part by the payment of page charges. Section solely to indicate this fact. You'll be in good company.

303 tonight 38 23901 38 of Lipid Research. Greeley Jr.

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Townsend Jr. Previous Section Next Section. Electrophysiology Cells growing on coverslips were mounted in a Leiden chamber and positioned on the stage of an inverted Nikon Diaphot phase contrast microscope. View this table: In this window In a new window.

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View larger version: Figure 3 Concentration response and time course of BBS-induced secretion. Previous Section. Marki W.

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Medline Google Scholar. Ladenheim E. CrossRef Medline Google Scholar.

Kim J. Cell Mol. Poitras P.

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G4 — G9. Kohjitani A. Weigert N. Kovacs T. G54 — G Cuttitta F. Bold R. Kroog G. Google Scholar. Kim H.

Aprikian A. Nemoz-Gaillard E. Evers B.

Parekh D. Jeng Y. Tsien R. Zhang T. Doyle H. Childs G.

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