Abstract
After permeabilization with the pore-forming toxin streptolysin-O mast cells can be triggered to secrete by addition of both calcium and a GTP analogue. If stimulation is delayed after permeabilization, there is a progressive decrease in the extent of secretion upon stimulation, eventually leading to a complete loss of the secretory response. This loss of secretory response can be retarded by the addition of cytosol from other secretory tissues, demonstrating that the response is dependent on a number of cytosolic proteins. We have used this as the basis of a bioassay to purify Secernin 1, a novel 50-kDa cytosolic protein that appears to be involved in the regulation of exocytosis from peritoneal mast cells. Secernin 1 increases both the extent of secretion and increases the sensitivity of mast cells to stimulation with calcium.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Calcium / pharmacology
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Cattle
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Chromatography
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Chromatography, Gel / methods*
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Cytosol / metabolism*
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Dose-Response Relationship, Drug
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Durapatite / pharmacology
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Exocytosis
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Glutathione Transferase / metabolism
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Humans
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Male
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Mass Spectrometry
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Mast Cells / metabolism*
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Molecular Sequence Data
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Nerve Tissue Proteins / chemistry*
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Nerve Tissue Proteins / isolation & purification*
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Plasmids / metabolism
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Proteins / chemistry*
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Proteins / isolation & purification*
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Rats
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Rats, Sprague-Dawley
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Recombinant Fusion Proteins / metabolism
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Sepharose / analogs & derivatives*
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Sepharose / pharmacology
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Sequence Homology, Amino Acid
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Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
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Time Factors
Substances
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Nerve Tissue Proteins
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Proteins
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Recombinant Fusion Proteins
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SCRN1 protein, human
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phenyl-superose
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secernin 1 protein, mouse
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Sepharose
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Durapatite
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Superose 12
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Glutathione Transferase
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Calcium