scholarly journals Chinese Hamster Ovary Cells Expressing a Cell Surface-anchored Form of Hepatic Lipase

1996 ◽  
Vol 271 (28) ◽  
pp. 16906-16914 ◽  
Author(s):  
Michael Komaromy ◽  
Salman Azhar ◽  
Allen D. Cooper
Keyword(s):  
Cell Surface ◽  
Chinese Hamster Ovary ◽  
Hepatic Lipase ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Ovary Cells ◽  
A Cell

scholarly journals Functional expression of the human transferrin receptor cDNA in Chinese hamster ovary cells deficient in endogenous transferrin receptor.

1987 ◽  
Vol 105 (1) ◽  
pp. 207-214 ◽  
Author(s):  
T E McGraw ◽  
L Greenfield ◽  
F R Maxfield
Keyword(s):  
Cell Surface ◽  
Chinese Hamster Ovary ◽  
Transferrin Receptor ◽  
Diphtheria Toxin ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Functional Expression ◽  
Genetically Engineered ◽  
Ovary Cell ◽  
Ovary Cells

Transferrin (Tf) receptor-variant Chinese hamster ovary cells have been isolated by selection for resistance to two Tf-toxin conjugates. The hybrid toxins contain Tf covalently linked to ricin A chain or a genetically engineered diphtheria toxin fragment. The Tf-receptor-variant (TRV) cells do not have detectable cell-surface Tf receptor; they do not bind fluorescein-Tf or 125I-Tf. TRV cells are at least 100-fold more resistant to the Tf-diphtheria toxin conjugate than are the parent cells. The TRV cells have retained sensitivity to native diphtheria toxin, indicating that the increased resistance to the conjugate is correlated with the loss of Tf binding. The endocytosis of fluorescein-labeled alpha 2-macroglobulin is normal in TRV cells, demonstrating that the defect does not pleiotropically affect endocytosis. Since these cells lack endogenous Tf receptor activity, they are ideally suited for studies of the functional expression of normal or altered Tf receptors introduced into the cells by cDNA transfection. One advantage of this system is that Tf binding and uptake can be used to monitor the behavior of the transfected receptor. A cDNA clone of the human Tf receptor has been transfected into TRV cells. In the stably expressing transfectants, the behavior of the human receptor is very similar to that of the endogenous Chinese hamster ovary cell Tf receptor. Tf binds to cell surface receptors, and is internalized into the para-Golgi region of the cell. Iron is released from Tf, and the apo-Tf and its receptor are recycled back to the cell surface. Thus, the TRV cells can be used to study the behavior of genetically altered Tf receptors in the absence of interfering effects from endogenous receptors.

Surface polypeptides of the Chinese hamster ovary cell; an immunochemical study

1976 ◽  
Vol 54 (2) ◽  
pp. 185-191 ◽  
Author(s):  
M. Behar-Bannelier ◽  
R. L. Juliano
Keyword(s):  
Cell Surface ◽  
Chinese Hamster Ovary Cell ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Ovary Cell ◽  
Immunochemical Study ◽  
Ovary Cells ◽  
Molecular Weights ◽  
Label Technique

Antibodies elicited by the injection of live Chinese hamster ovary cells (CHO) into rabbits precipitated four major components from detergent extracts of CHO membranes. The four components, of molecular weights 200 000, 125 000, 95 000 and 41 000 daltons, corresponded to cell surface components identified by the lactoperoxidase surface label technique.

scholarly journals Rapid cell surface appearance of endocytic membrane proteins in Chinese hamster ovary cells.

1981 ◽  
Vol 1 (3) ◽  
pp. 261-268 ◽  
Author(s):  
B Storrie ◽  
T D Dreesen ◽  
K M Maurey
Keyword(s):  
Membrane Proteins ◽  
Cell Surface ◽  
Chinese Hamster Ovary ◽  
Cho Cells ◽  
Gel Electrophoresis ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Surface Appearance ◽  
Ovary Cells ◽  
One Dimensional

Lactoperoxidase was used to selectively radiolabel endocytic membrane. CHO cells were incubated with enzyme at 37 degrees C for 10 min to permit lactoperoxidase internalization. Radioiodination was done at 4 degrees C. About 90% of the radioiodinated products pelleted at 100,000 X g. From 12 to 15 different electrophoretic species were detected by one-dimensional gel electrophoresis. When cells labeled by internalized lactoperoxidase were warmed to 37 degrees C, the incorporated radioactivity was lost in a biphasic manner with an overall t1/2 of approximately 20 h. Upon warming cells to 37 degrees C, the labeled species became sensitive to pronase or trypsin digestion. The increase in protease sensitivity was progressive over a 10- to 20-min period. Maximally 45% of the initially intracellular radiolabel could be released. A digest of exterior-radioiodinated cells released 50% of the incorporated radioiodine. These observations strongly suggest a rapid shuttling of approximately 90% of the radioiodinated membrane species initially present within the cell to the cell surface.

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