Co-localization of ras gene products and cathepsin L in cytoplasmic vesicles in v-Ha-ras-transformed NIH3T3 mouse fibroblasts.
Subcellular localisation of Ha-ras gene products (p21) and cathepsin L was investigated by immunocytochemical studies. Western blotting analysis by using v-Ha-ras p21 deletion-insertion mutants indicated that NCC-RAS-001 and -004 monoclonal antibodies recognize epitopes within amino acids 126-140 and 146-165, respectively, of Ha-ras p21. Indirect immunofluorescent studies using these antibodies showed that ras p21 is distributed in patch-shaped structures in the cytoplasm. Cathepsin L whose activity is effectively inhibited by p21s in vitro was found in the regions where p21s are concentrated. These p21-concentrated regions may be derived from endosome-like vesicles because they are also labeled with Lucifer Yellow. The co-localization of p21 and cathepsin L in cytoplasmic vesicles was also confirmed by electron microscopic immunocytochemistry.
Hiwasa
T
T
Division of Biochemistry, Chiba Cancer Center Research Institute, Japan.
Sawada
T
T
Tanaka
K
K
Chiba
T
T
Tanaka
T
T
Kominami
E
E
Katunuma
N
N
Sakiyama
S
S
D016475:3T3 Cells; D000818:Animals; D056668:Cathepsin L; D002403:Cathepsins / Q000378:metabolism*; D002471:Cell Transformation, Neoplastic; D003546:Cysteine Endopeptidases; D003593:Cytoplasm / Q000378:metabolism; D010450:Endopeptidases; D011905:Genes, ras; D007150:Immunohistochemistry; D051379:Mice; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics / Q000378:metabolism*
D002403:Cathepsins; D010450:Endopeptidases; D003546:Cysteine Endopeptidases; D056668:Cathepsin L; C534278:Ctsl protein, mouse; D016283:Proto-Oncogene Proteins p21(ras)