Determination of basal and stimulated levels of inositol triphosphate in (/sup 32/P)orthophosphate-labeled platelets
Journal Article
·
· Anal. Biochem.; (United States)
Previous studies indicated that thrombin-stimulation of platelets prelabeled with (/sup 3/H)inositol or (/sup 32/P)orthophosphate results in an increase of radioactive inositol triphosphate, a substance thought to modulate the levels of free intracellular calcium. In the present study, we improved the method of resolution of inositol triphosphate from other compounds that are also labeled with (/sup 32/P)orthophosphate using a combination of enzyme treatment and electrophoresis. We have further demonstrated that the specific activities of metabolic ATP and phosphatidylinositol diphosphate (the precursor of inositol triphosphate) are identical in (/sup 32/P)orthophosphate-labeled platelets. It follows that the amount of inositol triphosphate is proportional to its radioactivity in the metabolic compartment of the cells. Using this protocol, the concentration of inositol triphosphate in resting and thrombin-stimulated platelets were determined to be 1-4 and 10-30 pmol/10/sup 8/ cells, respectively.
- Research Organization:
- Temple Univ., Philadelphia, PA
- OSTI ID:
- 5578678
- Journal Information:
- Anal. Biochem.; (United States), Journal Name: Anal. Biochem.; (United States) Vol. 154:2; ISSN ANBCA
- Country of Publication:
- United States
- Language:
- English
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OSTI ID:6332649
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Thesis/Dissertation
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Tue Dec 31 23:00:00 EST 1985
·
OSTI ID:6974541
Related Subjects
550501* -- Metabolism-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ATP
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BLOOD PLATELETS
BODY FLUIDS
CARBOHYDRATES
COAGULANTS
DATA
DAYS LIVING RADIOISOTOPES
DRUGS
ENZYMES
ESTERS
EXPERIMENTAL DATA
HEMATOLOGIC AGENTS
HEMOSTATICS
HYDROLASES
INFORMATION
INOSITOLS
ISOTOPE APPLICATIONS
ISOTOPES
LABELLED COMPOUNDS
LABELLING
LIGHT NUCLEI
MATERIALS
MONOSACCHARIDES
NUCLEI
NUCLEOTIDES
NUMERICAL DATA
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
PEPTIDE HYDROLASES
PHOSPHORIC ACID ESTERS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PRECURSOR
QUANTITY RATIO
RADIOISOTOPES
SACCHARIDES
SERINE PROTEINASES
STIMULATION
THROMBIN
TRACER TECHNIQUES
59 BASIC BIOLOGICAL SCIENCES
ATP
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BLOOD PLATELETS
BODY FLUIDS
CARBOHYDRATES
COAGULANTS
DATA
DAYS LIVING RADIOISOTOPES
DRUGS
ENZYMES
ESTERS
EXPERIMENTAL DATA
HEMATOLOGIC AGENTS
HEMOSTATICS
HYDROLASES
INFORMATION
INOSITOLS
ISOTOPE APPLICATIONS
ISOTOPES
LABELLED COMPOUNDS
LABELLING
LIGHT NUCLEI
MATERIALS
MONOSACCHARIDES
NUCLEI
NUCLEOTIDES
NUMERICAL DATA
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
PEPTIDE HYDROLASES
PHOSPHORIC ACID ESTERS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PRECURSOR
QUANTITY RATIO
RADIOISOTOPES
SACCHARIDES
SERINE PROTEINASES
STIMULATION
THROMBIN
TRACER TECHNIQUES