z-logo
Premium
Intracellular distribution of active and inactive transglutaminase in stimulated cultured C 6 glioma cells
Author(s) -
Korner G.,
Bachrach U.
Publication year - 1987
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1041300108
Subject(s) - cycloheximide , stimulation , trypsin , tissue transglutaminase , enzyme , centrifugation , biochemistry , antigen , sendai virus , intracellular , specific activity , enzyme assay , microbiology and biotechnology , chemistry , differential centrifugation , biology , protein biosynthesis , immunology , endocrinology , gene
The cellular distribution of active and inactive transglutaminase (TGase) was studied in C 6 glioma cells before and during stimulation by a serum‐containing medium. The activity of the enzyme was determined in the soluble and insoluble fractions obtained by freezing and thawing the cells, followed by centrifugation at 12,000g for 5 min. In the soluble fractions, the activity of TGase decreased 2.5 h post‐stimulation and increased after 5 and 8 h. In the corresponding insoluble fractions, no significant changes in the activity of the enzyme were noted up to 8 h after stimulating the cells with fresh medium. An immunological approach was next used to determine the quantity of TGase antigen during the stimulation of the cultured glioma cells. In the soluble fraction, the quantity of the antigen decreases significantly at 2.5, 5, and 8 h. In contrast, in the insoluble fraction, a significant increase in TGase antigen was detercted 8 h after the addition of fresh medium. Cycloheximide completely inhibited the increase in the quantity of TGase antigen in the insoluble fraction, 8 h post‐stimulation, while actinomycin D caused a partial inhibition. Trypsin, neuraminidase, or Sendai viruses increased the activity of TGase significantly, when added to nonstimulated cells. Trypsin had no effect on TGase activity when added to the cells 2 h after stimulation with a serumcontaining medium. These findings suggest that an inactive form of the enzyme is present in the insoluble cellular fraction. A model has been proposed to explain the variations in TGase activity, its distribution and translocation during cellular stimulation.

This content is not available in your region!

Continue researching here.

Having issues? You can contact us here