Isolation and characterization of endosomes from rat liver
Three fractions of rat liver endosomes, called 50 Kg Light, 50 Kg Heavy, and 150 Kg have been isolated on 16% Percoll gradients. The 50 Kg Heavy fraction accumulates ligand as a function of time after injection, using either /sup 125/I-asialoorosomucoid (/sup 125/I-ASOR) or /sup 125/I-immunoglobulin A (/sup 125/I-IgA) as ligands. A pulse-chase protocol was also used to study the kinetics of ligand entry into the endosomal compartments. A double-label, 3,3'-diaminobenzidine (DAB)-induced density shift protocol was used to study the internalization of two ligands with different destinations in the hepatocyte. Rats were injected intraportally with /sup 125/I-ASOR-HRP and /sup 131/I-IgA and the liver was fractionated at various times post-injection. The three ligand-containing endosomal fractions were isolated and each subjected to the DAB shift procedure. This treatment causes organelles containing /sup 125/I-ASOR-HRP and another ligand occupying the same compartment to shift to a higher density. Thus, information on whether the /sup 131/I-IgA is colocalized or segregated from the /sup 125/I-ASOR-HRP can be obtained. The authors have used an instantaneous pulse, temperature shift protocol to study the heterogeneity of these three endosomal fractions isolated from rat liver.
- Research Organization:
- Case Western Reserve Univ., Cleveland, OH (USA)
- OSTI ID:
- 6905389
- Resource Relation:
- Other Information: Thesis (Ph. D.)
- Country of Publication:
- United States
- Language:
- English
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LABELLED COMPOUNDS
UPTAKE
LIGANDS
LIVER CELLS
BIOCHEMICAL REACTION KINETICS
COMPARTMENTS
IMMUNOGLOBULINS
IODINE 125
IODINE 131
LIVER
MUCOPROTEINS
RATS
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CARBOHYDRATES
DAYS LIVING RADIOISOTOPES
DIGESTIVE SYSTEM
ELECTRON CAPTURE RADIOISOTOPES
GLANDS
GLOBULINS
INTERMEDIATE MASS NUCLEI
IODINE ISOTOPES
ISOTOPES
KINETICS
MAMMALS
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANS
POLYSACCHARIDES
PROTEINS
RADIOISOTOPES
REACTION KINETICS
RODENTS
SACCHARIDES
SOMATIC CELLS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques