scholarly journals Expression and maturation of human cathepsin D in baby-hamster kidney cells

1991 ◽  
Vol 273 (2) ◽  
pp. 355-361 ◽  
Author(s):  
M Horst ◽  
A Hasilik
Keyword(s):  
Cathepsin D ◽  
Human Fibroblasts ◽  
Apparent Size ◽  
Human Cells ◽  
Kidney Cells ◽  
Baby Hamster Kidney ◽  
Transfected Cells ◽  
Elevated Activity ◽  
Human Cathepsin ◽  
Baby Hamster Kidney Cells

In medium and in homogenates from baby-hamster kidney cells (BHK) transfected with human cathepsin D cDNA, an elevated activity of cathepsin D was found as compared to non-transfected cells. The elevated activity was removed by titrating the homogenates with an anti-(human cathepsin D) antibody. Metabolic labelling and immunoprecipitation revealed that, in the transfected cells, human cathepsin D was synthesized as a 53-kDa precursor indistinguishable from that found in human cells. A portion of the precursor was secreted and the remainder was processed to intermediate and mature chains within a few hours of synthesis. The precursor that was released from the transfected cells had a slightly smaller apparent size than that from cultured human fibroblasts. This difference was abrogated when the precursors were treated with glycopeptidase F. In the intracellular small chain a difference was observed in the size of carbohydrate chains that were cleavable with endo-beta-N-acetylglucosaminidase H. Sequence analysis of the N-termini of mature intracellular cathepsin D indicated a N-terminal trimming in both large and small chains from both human and transfected hamster cells. The proteolytic maturation of human cathepsin D in BHK cells closely resembles that in human cells, whereas a portion of the carbohydrate side chains is processed differently. The trimming of the N-termini in mature cathepsin D is proposed to be a part of the maturation and aging of this protein.

scholarly journals Differential segregation of human and hamster cathepsin D in transfected baby-hàmster kidney cells

1991 ◽  
Vol 273 (2) ◽  
pp. 363-367 ◽  
Author(s):  
C Isidoro ◽  
M Horst ◽  
F M Baccino ◽  
A Hasilik
Keyword(s):  
Cathepsin D ◽  
High Rate ◽  
Kidney Cells ◽  
Baby Hamster Kidney ◽  
Human Enzyme ◽  
Before And After ◽  
Human Cathepsin ◽  
Low Rate ◽  
Baby Hamster Kidney Cells ◽  
In Cells

The segregation of human cathepsin D, studied in baby-hamster kidney cells (BHK) transfected with human cathepsin D cDNA and compared with that of hamster cathepsin D in the same cells, showed that, in cells that expressed human cathepsin D at a low rate, most of the enzyme remained intracellular. In contrast, when the enzyme was expressed at a high rate, most was secreted. The segregation was examined with an anti-(human cathepsin D) antibody that reacted with the human enzyme exclusively and an anti-(rat cathepsin D) antibody that reacted with both enzymes. In one protocol the cells were metabolically labelled and the two antibodies were used in sequence to precipitate the enzymes from extracts of cells and medium. High expression of the human enzyme did not interfere with the segregation of hamster cathepsin D. In another protocol the activity of cathepsin D in cells and medium was measured before and after titration with anti-(human cathepsin D) antiserum. Human cathepsin D was found predominantly in the medium, and hamster cathepsin D mainly in the cells. In the presence of 10 mM-NH4Cl the intracellular segregation of hamster cathepsin D was strongly inhibited, while the segregation of human cathepsin D was only slightly diminished. In BHK cells, at least two systems participate in the sorting of the two cathepsins, one of them being rather insensitive to NH4Cl.

Turbomolecular Pumped Electron Microscopy of Rubella Virus (In BHK Celis)

1968 ◽  
Vol 26 ◽  
pp. 204-205
Author(s):  
A. B. Taylor ◽  
G. C. Cole ◽  
M. A. Holcomb ◽  
C. A. Baechler
Keyword(s):  
Electron Microscopy ◽  
Rubella Virus ◽  
Phosphate Buffer ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Basal Medium ◽  
Kidney Cells ◽  
Baby Hamster Kidney ◽  
Input Multiplicity ◽  
Baby Hamster Kidney Cells

An aliquot from a continuous fermenter culture of baby hamster kidney cells (BHK-21 Clone PD-4) (Wistar) maintained in Ca free Eagle's Basal Medium containing 2% Kaolin adsorbed fetal calf serum was planted in spinner flasks at 300,000 cells per ml, total volume 600 ml. After equilibration for one day at 35°C to insure that cells were in log phase, the culture was infected with the M-33-AGMK25 BHK-219 strain of rubella at an input multiplicity of about 6 TCID50 per cell. The virus was identified with specific rubella antiserum.Preliminary experiments had shown that such cultures would reach a peak or plateau HA titer of approximately 1:64, 24 hrs after inoculation and would continue to yield virus for 6 to 12 days. One hundred ml aliquot harvests were withdrawn daily and the culture was returned to volume with growth medium and incubation continued. The harvested cells were spun down rapidly at 2500 rpm per 15 mins., fixed in 3.7% gluteraldehyde in Ca free phosphate buffer saline, and post fixed in osmium tetraoxide. After dehydration, the cells were embedded in Epon 812 and cured approximately 20 hrs at 60°C.

scholarly journals Allicin from garlic strongly inhibits cysteine proteinases and cytopathic effects of Entamoeba histolytica.

1997 ◽  
Vol 41 (10) ◽  
pp. 2286-2288 ◽  
Author(s):  
S Ankri ◽  
T Miron ◽  
A Rabinkov ◽  
M Wilchek ◽  
D Mirelman
Keyword(s):  
Alcohol Dehydrogenase ◽  
Entamoeba Histolytica ◽  
Kidney Cells ◽  
Cysteine Proteinases ◽  
Baby Hamster Kidney ◽  
Important Contributor ◽  
Cytopathic Effects ◽  
Baby Hamster Kidney Cells

The ability of Entamoeba histolytica trophozoites to destroy monolayers of baby hamster kidney cells is inhibited by allicin, one of the active principles of garlic. Cysteine proteinases, an important contributor to amebic virulence, as well as alcohol dehydrogenase, are strongly inhibited by allicin.

scholarly journals Cytotoxicity of Pristine C<sub>60</sub> Fullerene on Baby Hamster Kidney Cells in Solution

2012 ◽  
Vol 03 (03) ◽  
pp. 385-390 ◽  
Author(s):  
Shufang Liu ◽  
Haijie Liu ◽  
Zhijuan Yin ◽  
Kai Guo ◽  
Xibao Gao
Keyword(s):  
Kidney Cells ◽  
Baby Hamster Kidney ◽  
Baby Hamster Kidney Cells

scholarly journals Uptake and utilization of 5'-methylthioadenosine by cultured baby-hamster kidney cells

1982 ◽  
Vol 204 (3) ◽  
pp. 803-807 ◽  
Author(s):  
T O Eloranta ◽  
K Tuomi ◽  
A M Raina
Keyword(s):  
Nucleic Acids ◽  
Cellular Metabolism ◽  
Exponential Phase ◽  
Kidney Cells ◽  
Baby Hamster Kidney ◽  
Purine Nucleotide ◽  
Salvage Pathway ◽  
Baby Hamster Kidney Cells

5'-Methylthioadenosine was taken up and immediately metabolized further by cultured baby-hamster kidney cells during the exponential phase of growth. The adenine moiety supplied the purine-nucleotide pool via the salvage pathway and was efficiently incorporated into nucleic acids. Catabolites of methylthioadenosine excreted by the cells included adenine, purinic compounds and metabolites of the ribose portion. 5'-Methylthiotubercidin had no significant effect on the cellular metabolism of methyl-thioadenosine, but greatly inhibited its uptake. erythro-9-(2-Hydroxy-3-nonyl)adenine had no effect on the uptake, but markedly interfered with the further utilization of methylthioadenosine after cleavage in the cells.

scholarly journals Clonal growth of hamster free alveolar cells in soft agar.

1975 ◽  
Vol 142 (4) ◽  
pp. 877-886 ◽  
Author(s):  
H S Lin ◽  
C Kuhn ◽  
T Kuo
Keyword(s):  
Clonal Growth ◽  
Soft Agar ◽  
Culture Conditions ◽  
Mononuclear Phagocytes ◽  
Kidney Cells ◽  
Baby Hamster Kidney ◽  
Alveolar Cells ◽  
Bronchial Washing ◽  
Lag Period ◽  
Baby Hamster Kidney Cells

Free alveolar cells obtained from healthy unstimulated hamsters were tested for their ability to form colonies in soft agar. Every bronchial washing so far tested contained colon-forming cells. The average plating efficiency was 8.1% (2.4-18.3%). Alveolar colony-forming cells were characterized by having a long initial lag period (4-8 days) and only mononuclear phagocytes were found in the colony. Medium conditioned by baby hamster kidney cells or other cells was required for the initiation and maintenance of their growth. Alveolar cells from normal mice and rats also formed colonies under appropriate culture conditions.

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