Pepstatin is a pentapeptide, a naturally occurring aspartyl protease inhibitor, which can inhibit aspartic protease and acid protease of various microorganisms. Pepstatin is mainly secreted by Streptomyces species and produced by Streptomyces. It can inhibit pepsin, pepsin D and angiotensin-releasing enzyme, and has therapeutic effects on gastric ulcer, renal hypertension, arthritis, carrageenan edema and other diseases.
Pepstatin is a strong inhibitor of aspartyl proteases such as pepsin, cathepsin D, and renin. This natural pentapeptide isolated from actinomycetes was for many years the classic renin inhibitor in vitro.Pepstatin is not specific for renin and is poorly soluble in water. Structural derivatives of pepstatin increased its solubility and specificity for renin by several orders of magnitude.Pepstatin contains the unusual γ amino acid statin that may substitute for the two amino acids at the scissile bond of the protein substrate and block substrate cleavage because of the structural analogy to a transition state of the peptide bond hydrolysis by aspartyl proteases.
When protein is extracted from broken cells, proteases can be released, which need to be rapidly inhibited to keep protein from being degraded. In the process of protein extraction, protease inhibitors need to be added to prevent proteolysis. Protease inhibitor broadly refers to a substance that binds to some groups on the active center of protease molecules, so that the protease activity decreases or even disappears, but does not denature the enzyme protein. The sensitivity of various proteases to different protein is different, so it is necessary to adjust the concentration of various proteases. Because the solubility of protease inhibitor in liquid is extremely low, it should be paid special attention to that protease inhibitor should be fully mixed in the buffer to reduce the precipitation of protease inhibitor. Pepstantin A can inhibit acid proteases such as pepsin, angiotensin, collagenase, cathepsin D and chymosin.
Pepstatin A is an inhibitor of cathepsin d and e. After HEK293 cells were treated with different concentrations of pepstatin A for 24 hours, the expression of LC3Ⅱ and p62 was detected. The results showed that pepstatin A could significantly enhance the expression of LC3Ⅱ and P62 (P < 0.05) in a dose-dependent manner. 20μ g/ml pepstatin A was used to treat HEK293 at different time intervals, and the effects of different time intervals on the expression of LC3II and p62 were observed. The results showed that pepstatin A could up-regulate the expression of LC3II and p62 in a time-dependent manner.
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