IMMUNOLOCALIZATION OF THIMET OLIGOPEPTIDASE IN CHICKEN EMBRYONIC FIBROBLASTS

被引:7
作者
CHEN, JM [1 ]
CHANGCO, A [1 ]
BROWN, MA [1 ]
BARRETT, AJ [1 ]
机构
[1] THOMAS JEFFERSON UNIV,JEFFERSON MED COLL,DEPT ANAT & DEV BIOL,PHILADELPHIA,PA 19107
关键词
D O I
10.1006/excr.1995.1010
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
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. (C) 1995 Academic Press, Inc.
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页码:80 / 85
页数:6
相关论文
共 47 条
[1]   SYNAPTOSOMAL MEMBRANE-BOUND FORM OF ENDOPEPTIDASE-24.15 GENERATES LEU-ENKEPHALIN FROM DYNORPHIN1-8, ALPHA-NEUENDORPHIN-BETA-NEOENDORPHIN, AND MET-ENKEPHALIN FROM MET-ENKEPHALIN-ARG6-GLY7-LEU8 [J].
ACKER, GR ;
MOLINEAUX, C ;
ORLOWSKI, M .
JOURNAL OF NEUROCHEMISTRY, 1987, 48 (01) :284-292
[2]  
ALLISON AC, 1969, LYSOSOM BIOL PATHOL, V2, P601
[3]  
BACKER JM, 1990, J BIOL CHEM, V265, P14828
[4]  
Barrett A J, 1992, Matrix Suppl, V1, P95
[5]   THE ACTIVITIES OF PZ-PEPTIDASE AND ENDOPEPTIDASE-24.15 ARE DUE TO A SINGLE ENZYME [J].
BARRETT, AJ ;
TISLJAR, U .
BIOCHEMICAL JOURNAL, 1989, 261 (03) :1047-1050
[6]   CHICKEN LIVER PZ-PEPTIDASE, A THIOL-DEPENDENT METALLOENDOPEPTIDASE [J].
BARRETT, AJ ;
BROWN, MA .
BIOCHEMICAL JOURNAL, 1990, 271 (03) :701-706
[7]   THIMET OLIGOPEPTIDASE (EC 3.4.24.15) - THE SAME BY ANY NAME [J].
BARRETT, AJ .
BIOCHEMICAL JOURNAL, 1991, 277 :295-296
[8]   A RAPID, SENSITIVE METHOD FOR DETECTION OF ALKALINE-PHOSPHATASE CONJUGATED ANTI-ANTIBODY ON WESTERN BLOTS [J].
BLAKE, MS ;
JOHNSTON, KH ;
RUSSELLJONES, GJ ;
GOTSCHLICH, EC .
ANALYTICAL BIOCHEMISTRY, 1984, 136 (01) :175-179
[9]  
BLUM JS, 1991, J BIOL CHEM, V266, P22091
[10]   ENDOCYTOSIS IN FILTER-GROWN MADIN-DARBY CANINE KIDNEY-CELLS [J].
BOMSEL, M ;
PRYDZ, K ;
PARTON, RG ;
GRUENBERG, J ;
SIMONS, K .
JOURNAL OF CELL BIOLOGY, 1989, 109 (06) :3243-3258