Efficient degradation in vitro of all intermediate filament subunit proteins by the Ca2+-activated neutral thiol proteinase from Ehrlich ascites tumor cells and porcine kidney.
نویسندگان
چکیده
Vimentin, desmin, glial fibrillary acidic protein, neurofilament triplet proteins, and a mixture of cytokeratins were digested with Ca2+-activated neutral thiol proteinase isolated from Ehrlich ascites tumor (EAT) cells and porcine kidney. All intermediate filament proteins were degraded by the proteinase, although with different rates and Ca2+ optima. These results are in part at variance with our previous statement that the Ca2+-activated proteinase from EAT cells is specific for vimentin and desmin.
منابع مشابه
Purification and further characterization of the Ca2+-activated proteinase specific for the intermediate filament proteins vimentin and desmin.
A calcium (Ca2+)-activated, neutral proteinase has been purified from Ehrlich ascites tumor cells. The protocol used has resulted in a 3,600-fold purification of the enzyme in a yield of 21% from the Ehrlich ascites tumor cell postnuclear supernatant. The purified proteinase has a high substrate specificity for the intermediate filament subunit proteins, vimentin and desmin, and showed no activ...
متن کاملIntermediate (10 nm) filament proteins and the Ca2+-activated proteinase specific for vimentin and desmin in the cells from fish to man: an example of evolutionary conservation.
Cultured cells from representative species of the five main Classes of Chordates (Mammalia, Aves, Reptilia, Amphibia and Pisces) were examined for the presence of intermediate (10 nm) filament proteins. Using a combination of morphological and immunological methods and detailed protein-chemical characterization it is demonstrated that intermediate filament proteins, as shown also for actin and ...
متن کاملPurification of the intermediate filament protein vimentin from Ehrlich ascites tumor cells.
Two procedures have been used for the purification of the intermediate filament protein vimentin from Ehrlich ascites tumor (EAT) cells grown in vitro. In one procedure, vimentin was first incorporated into residual cell structures by extracting EAT cells with Triton X-100 in the presence of M g ’ and then solubilized with low ionic strength buffer in the absence of M g + . In the second proced...
متن کاملInvolvement of the N-terminal polypeptide of vimentin in the formation of intermediate filaments.
The potential to form intermediate filaments of a 54 X 10(3) molecular weight (Mr) polypeptide derived from vimentin by cleavage by the intermediate filament-specific, Ca2+-activated proteinase was investigated. Under physiological conditions of assembly, the breakdown product did not form intermediate filaments. Electron microscopy revealed short, rod-like structures similar to those described...
متن کاملPolyacrylamide gel electrophoretic screening of mammalian cells cultured in vitro for the presence of the intermediate filament protein vimentin.
A total of 53 different cell lines originating from a variety of mammalian species were cultured in vitro and analysed for the presence of vimentin, employing polyacrylamide gradient slab gel electrophoresis in urea/acetic acid as buffer system. Irrespective of the cell culture conditions, and the growth potential and morphology of the cells, vimentin was expressed in all cell lines examined, w...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Bioscience reports
دوره 6 1 شماره
صفحات -
تاریخ انتشار 1986