Immunolocalization of thimet oligopeptidase in chicken embryonic fibroblasts. 1995

J M Chen, and A Changco, and M A Brown, and A J Barrett
Department of Anatomy and Developmental Biology, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania 19107.

An antiserum was raised against chicken thimet oligopeptidase (TOP), and specific antibodies were isolated by immunoadsorption. The specificity of the antibodies was confirmed by immunoblot analysis with the cell extract of cultured chicken fibroblasts. Subcellular localization of TOP in normal and Rous sarcoma virus-transformed chicken embryonic fibroblasts was investigated by indirect immunofluorescence staining with affinity-purified, monospecific anti-TOP antibodies. In addition to the diffuse reaction expected for a cytosolic enzyme, a punctate, cytoplasmic localization was observed. The appearance of the organelles containing TOP did not correspond to that expected for secondary lysosomes, mitochondria, endoplasmic reticulum, or Golgi apparatus, but did resemble that of endosomes. When cells were allowed to internalize Lucifer yellow before fixation, some of the organelles containing TOP were observed to contain Lucifer yellow. Moreover, those organelles containing TOP were positively immunolabeled with the anti-transferrin receptor antibody. We conclude that part of the TOP in chicken embryonic fibroblasts is located in endosomes and discuss the implications of this.

UI MeSH Term Description Entries
D007546 Isoquinolines A group of compounds with the heterocyclic ring structure of benzo(c)pyridine. The ring structure is characteristic of the group of opium alkaloids such as papaverine. (From Stedman, 25th ed)
D008666 Metalloendopeptidases ENDOPEPTIDASES which use a metal such as ZINC in the catalytic mechanism. Metallo-Endoproteinases,Metalloendopeptidase
D011990 Receptors, Transferrin Membrane glycoproteins found in high concentrations on iron-utilizing cells. They specifically bind iron-bearing transferrin, are endocytosed with its ligand and then returned to the cell surface where transferrin without its iron is released. Transferrin Receptors,Transferrin Receptor,Receptor, Transferrin
D011992 Endosomes Cytoplasmic vesicles formed when COATED VESICLES shed their CLATHRIN coat. Endosomes internalize macromolecules bound by receptors on the cell surface. Receptosomes,Endosome,Receptosome
D002642 Chick Embryo The developmental entity of a fertilized chicken egg (ZYGOTE). The developmental process begins about 24 h before the egg is laid at the BLASTODISC, a small whitish spot on the surface of the EGG YOLK. After 21 days of incubation, the embryo is fully developed before hatching. Embryo, Chick,Chick Embryos,Embryos, Chick
D005347 Fibroblasts Connective tissue cells which secrete an extracellular matrix rich in collagen and other macromolecules. Fibroblast
D005456 Fluorescent Dyes Chemicals that emit light after excitation by light. The wave length of the emitted light is usually longer than that of the incident light. Fluorochromes are substances that cause fluorescence in other substances, i.e., dyes used to mark or label other compounds with fluorescent tags. Flourescent Agent,Fluorescent Dye,Fluorescent Probe,Fluorescent Probes,Fluorochrome,Fluorochromes,Fluorogenic Substrates,Fluorescence Agents,Fluorescent Agents,Fluorogenic Substrate,Agents, Fluorescence,Agents, Fluorescent,Dyes, Fluorescent,Probes, Fluorescent,Substrates, Fluorogenic
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D000906 Antibodies Immunoglobulin molecules having a specific amino acid sequence by virtue of which they interact only with the ANTIGEN (or a very similar shape) that induced their synthesis in cells of the lymphoid series (especially PLASMA CELLS).
D000918 Antibody Specificity The property of antibodies which enables them to react with some ANTIGENIC DETERMINANTS and not with others. Specificity is dependent on chemical composition, physical forces, and molecular structure at the binding site. Antibody Specificities,Specificities, Antibody,Specificity, Antibody

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