Detection and assay of proteases using calf lens beta-crystallin aggregate as substrate.
The eye lens protein, betaL-crystallin, aggregates and yields a turbid solution upon refolding from its denatured state. We have observed that the addition of trace amounts of protease results in clearing of this turbidity. Based on this observation, we have developed a simple and rapid method for the detection and assay of proteases. This assay can be performed in the pH range of 6.0-9.0. We could assay the activity of trypsin at a concentration as low as 5 microg/ml.
Trivedi
V D
VD
Centre for Cellular and Molecular Biology, Hyderabad, India.
Raman
B
B
Ramakrishna
T
T
Rao
C M
CM
D000818:Animals; D002417:Cattle; D003459:Crystallins / Q000378:metabolism*; D010450:Endopeptidases / Q000032:analysis*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009391:Nephelometry and Turbidimetry / Q000379:methods; D011480:Protease Inhibitors / Q000494:pharmacology; D011489:Protein Denaturation; D017510:Protein Folding; D012995:Solubility; D014357:Trypsin / Q000032:analysis
D003459:Crystallins; D011480:Protease Inhibitors; D010450:Endopeptidases; D014357:Trypsin