Protease activities in cultured beef lens epithelial cells peak and then decline upon progressive passage.
Beef lens cells in culture are readily obtained and provide many opportunities to study phenomena related to cell differentiation and maturation, environmental stress, disease, and perhaps mechanisms of transformation. Although altered rates of proteolysis are known to accompany these phenomena, the proteolytic activities available in cultured beef lens epithelial cells have not been documented. In this work are documented the specific activities, based on protein and DNA content, of neutral exo- and endopeptidase, cathepsins B- and D-like enzymes and acid phosphatase in lens epithelial cortical and core tissue and in cultured epithelial cells at passages 1-43. Maximal activity of each protease occurs almost routinely at passage 5 or 9, reaching values of approx. 1400-, 0.77-, 4520-nmol min-1 per mg protein for neutral exopeptidase (passage 5), neutral endopeptidase (passage 5) and cathepsin B (passage 5) respectively, and 7.1 micrograms trichloroacetic acid soluble peptide min-1 per mg protein for cathepsin D (passage 15). On a microgram-1 DNA basis, the maximal specific activities for the same enzymes were 48 (passage 5), 0.03 (passage 5), 283 (passage 9), and 0.5 (passage 9) respectively. In subsequent passages, the specific activities declined to values which were similar to or lower than the specific activities observed for these proteases in lens epithelial tissue.
Eisenhauer
D A
DA
USDA Human Nutrition Research Center on Aging, Tufts University, Boston, MA 02111.
Berger
J J
JJ
Peltier
C Z
CZ
Taylor
A
A
D000135:Acid Phosphatase / Q000378:metabolism; D000818:Animals; D002401:Cathepsin B / Q000378:metabolism; D002402:Cathepsin D / Q000378:metabolism; D002417:Cattle; D002478:Cells, Cultured; D003459:Crystallins / Q000032:analysis; D004247:DNA / Q000032:analysis; D010450:Endopeptidases / Q000378:metabolism; D004848:Epithelium / Q000201:enzymology; D020689:Exopeptidases; D007908:Lens, Crystalline / Q000201:enzymology*; D015260:Neprilysin; D010447:Peptide Hydrolases / Q000378:metabolism*; D013997:Time Factors
D003459:Crystallins; D004247:DNA; D000135:Acid Phosphatase; D010450:Endopeptidases; D020689:Exopeptidases; D010447:Peptide Hydrolases; D002401:Cathepsin B; D002402:Cathepsin D; D015260:Neprilysin