Effects of pH, detergent and salt on aggregation of Chinese-hamster-ovary-cell lysosomal enzymes.
نویسندگان
چکیده
Upon detergent or hypo-osmotic lysis of CHO-cell postnuclear supernatants or isolated lysosomes at pH 4.8, the lysosomal enzymes beta-hexosaminidase, beta-galactosidase, alpha-fucosidase and cathepsin C were readily pelleted, whereas the exogenous marker, long-term-internalized horseradish peroxidase, was not. Salt or pH elevation greatly decreased lysosomal-enzyme pelletability. The results suggest that, under native conditions, lysosomal hydrolases may be aggregated. Aggregation could promote enzyme retention within the organelle.
منابع مشابه
Propranolol induced chromosomal aberrations in Chinese hamster ovary cell line
Propranolol (PL), a non-selective beta-blocker, is a cardiovascular drug widely used to treat hypertension. The present study was concerned with assessing the cytogenetic effects of this drug on Chinese hamster ovary (CHO) cell line. MTT assay was then carried out to determine the cytotoxicity index (IC50) of the drug. The IC50 value of PL was 0.43±0.02 mM. To investigate the clastogenic effect...
متن کاملP-76: Stepwise Reduction of Fetal Bovine Serum Levels in Chinese Hamster OvaryCells -Expressing Human Chorionic Gonadotrophin- Culture
Background: The demand for producing recombinant therapeutic proteins by mammalian cell lines, such as Chinese hamster ovary (CHO) cells, continues to grow. Significant achievements in process optimization including development of cell culture strategies for largescale and cost-effective production have been made. Fetal bovine serum (FBS) is an often essential growth supplement and yet most cos...
متن کاملP-234: Expression of Human Chorionic Gonadotropin (hCG) Hormone Using Chinese Hamster Ovary Cells
Background: Human chorionic gonadotropin (hCG) is a member of glycoprotein hormones family consist of two different non-covalently heterodimeric chains: alpha and beta subunits with 92 and 145 amino acids respectively. This hormone plays an important role in human reproduction and physiology especially for maintenance of the corpus luteum during the first months of pregnancy Materials and Metho...
متن کاملCloning and Expression of Recombinant Human Interleukin-7 in Chinese Hamster Ovary (CHO) Cells
Background: The critical role of interleukin-7 (IL-7) in homeostatic proliferation and T cell survival has made it a promising cytokine for the treatment of various clinical conditions, especially those associated with lymphopenia. Methods: In the present study we expressed recombinant human interleukin-7 (rhIL-7) in Chinese hamster ovary (CHO)-K1 cells. CHO-K1 cells were stably transfected ...
متن کاملIntracellular Localization of FLAG-Peroxisomal Protein in Chinese Hamster Ovary (CHO) Cells
Epitope tagging is a method of expressing proteins whereby an epitope for a specific monoclonal antibody is fused to a target protein using recombinant DNA techniques. The aim of this study was to sub-clone the peroxisomal protein (PEP) cDNA into a mammalian expression vector leading to the formation of a chimeric PEP-cDNA containing the FLAG epitope. The FLAG-PEP recombinant cDNA was construc...
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:
- The Biochemical journal
دوره 249 3 شماره
صفحات -
تاریخ انتشار 1988