Characteristics of pancreatic cholesterol esterase binding to and uptake by rat intestinal cells
Thesis/Dissertation
·
OSTI ID:5642787
In the intestinal lumen cholesterol esterase derived from pancreatic juice catalyzes the hydrolysis of cholesteryl esters (CE). The characteristics of Ce'ase binding to and uptake by rat intestinal cells were determined. CE'ase purified from rat pancreas with a specific activity 2 fold higher and a yield 5 fold greater than that previously attainable was judged as homogeneous on the basis of SDS-PAGE and sedimentation equilibrium centrifugation. Intestinal cell types and membranes were isolated and judged as pure on the basis of marker enzyme analyses. The enzyme was radiolabeled with ({sup 125}-I) to a specific radioactivity of 55 Ci/mmole with retention of biological activity, gross molecular size, secondary structure, and immunological properties. ({sup 125}-I) CE'ase bound preferentially to mature absorptive cells from proximal intestine and their brush border membranes. A specific, low affinity binding phenomenon was demonstrated with the following characteristics: linearity with increasing ligand concentration (non-saturability) or cell concentration, time and temperature dependency, and irreversibility. Native CE'ase, at a 500 fold molar excess did not displace bound ({sup 125}-I) CE'ase.
- Research Organization:
- George Washington Univ., Washington, DC (USA)
- OSTI ID:
- 5642787
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
BETA DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BODY
CELL CONSTITUENTS
CELL MEMBRANES
CENTRIFUGATION
CHEMICAL COMPOSITION
CHOLESTEROL
DAYS LIVING RADIOISOTOPES
DIGESTIVE SYSTEM
ELECTRON CAPTURE RADIOISOTOPES
ELECTROPHORESIS
ENDOCRINE GLANDS
ENZYMES
ESTERASES
GASTROINTESTINAL TRACT
GLANDS
HYDROLASES
HYDROXY COMPOUNDS
INTERMEDIATE MASS NUCLEI
INTESTINES
IODINE 125
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
MAMMALS
MEMBRANES
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANS
PANCREAS
RADIOISOTOPES
RATS
REACTION KINETICS
RODENTS
SEPARATION PROCESSES
STEROIDS
STEROLS
TEMPERATURE DEPENDENCE
TIME DEPENDENCE
TRACER TECHNIQUES
ULTRACENTRIFUGATION
VERTEBRATES
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
BETA DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BODY
CELL CONSTITUENTS
CELL MEMBRANES
CENTRIFUGATION
CHEMICAL COMPOSITION
CHOLESTEROL
DAYS LIVING RADIOISOTOPES
DIGESTIVE SYSTEM
ELECTRON CAPTURE RADIOISOTOPES
ELECTROPHORESIS
ENDOCRINE GLANDS
ENZYMES
ESTERASES
GASTROINTESTINAL TRACT
GLANDS
HYDROLASES
HYDROXY COMPOUNDS
INTERMEDIATE MASS NUCLEI
INTESTINES
IODINE 125
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
MAMMALS
MEMBRANES
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANS
PANCREAS
RADIOISOTOPES
RATS
REACTION KINETICS
RODENTS
SEPARATION PROCESSES
STEROIDS
STEROLS
TEMPERATURE DEPENDENCE
TIME DEPENDENCE
TRACER TECHNIQUES
ULTRACENTRIFUGATION
VERTEBRATES