No abstract available.
Neoplasm Metastasis* ; Peptide Hydrolases*

Proteolytic activity of enzyme in some preparations of granules, tablets and capsules were changed during the production processing. The decrease of proteolytic activity was dependent on the origin and purity of the enzymatic preparations
Enzymology ; Pharmaceutical Preparations ; Peptide Hydrolases

The reasonable pepsin + papain mix was studied. The results showed that the proteolytic activity of the mix at 3 areas pH: 20.5, 41 and 70.5 and temperature 38O2 was compared with either agent alone in the same conditions. These mixes will be improved and applied to the treatment of malnutrition in children
Peptide Hydrolases ; Enzymology ; Pepsin A ; Papain

A keratinolytic proteinase secreted by Microsporum canis in a broth containing human hair was purified 134-fold from the culture filtrate by ion-exchange chromatography using DEAE-Sephacel, CM-Sephadex C-50, and by Sephadex G-75 gel filtration. The purified enzyme was electrophoretically homogeneous with a molecular weight of 33,000. The enzyme had an optimum pH of 8.0, and the activity was stable in the alkaline pH range. Enzyme activity increased with temperature up to 35 degrees C and was stable up to 45 degrees C. The keratinolytic activity was not affected by the addition of nonionic detergents, was activated by Mg2+, but inhibited by Zn2+. The purified enzyme was used to obtain guinea pig antiserum. The antiserum tested by double diffusion against the purified enzyme showed a single line of precipitation and completely neutralized the proteinase activity. This study reaffirms that the proteinase from M. canis may be a biochemical mechanism for the invasion of keratinized tissue, and could possibly play a role in the hypersensitivity reactions arising from superficial infections of this fungus.
Microsporum/enzymology* ; Peptide Hydrolases/isolation & purification*

Viruses interact with the host ubiquitination system in a variety of ways. Viral proteins are often a substrate for ubiquitination, which leads to proteasomal degradation. Viruses also have functions to modify the cellular ubiquitination machinery. Recently, deubiquitinating protease (DUB) activity has been found in many viral proteins. In herpesviruses, the DUB domain is found within the large tegument protein, which is conserved in all members of the herpesvirus family. Although a limited number of viral and cellular targets have been identified to date, accumulating evidence shows that herpesviral DUBs may primarily target key cellular regulators of immune signaling pathways to promote viral replication. In this review, we summarize the recent findings on viral DUBs. In particular, we focus on the herpesviral DUBs and their targets, and discuss their potential roles in the regulation of immune signaling pathways.
Herpesviridae ; Humans ; Peptide Hydrolases* ; Ubiquitin ; Ubiquitination ; Viral Proteins

Serine elastic chymotrypsin Pr1 is an enzyme that efficiently degrades insect body wall protein through its connection with the virulence of entomogenous fungi. Therefore, it is important to explore the relationship between the Pr1 protease activity, the Pr1 gene expression and the virulence of different strains of entomogenous fungi. Specific peptide substrate Suc-Ala-Ala-Pro-Phe-pNA and fluorogenic quantitative PCR were used for detecting Pr1 protease activity and Pr1 gene expression, and the slope spray method was used for evaluating the virulence of the fungi on the Myzus persicae. The results indicated that the linear regression equation of the Pr1 protease activity and the virulence of different strains were: y=3.64x+0.62, R²=0.432. It was shown that there is a positive correlation between the Pr1 protease activity and virulence of different strains. Moreover, the result of the multiple linear regression analysis between Pr1 protease activity, Pr1 gene expression and the virulence of different strains was: y=0.236+10.833x₁-0.039x₂ (x₁ represents Pr1 protease activity while x₂ represents Pr1 gene expression), R²=0.568, which suggested that the raw data could be represented by a linear fitting equation. The serial correlation coefficient was high (D-W was 2.444), indicating that Pr1 protease activity and Pr1 gene expression have great effect on the virulence of the fungi. Additionally, VIF=12.705, which shows that moderate multiple collinear exists between Pr1 protease activity and Pr1 gene expression. Therefore, Pr1 protease activity and Pr1 gene expression could be recommended as important indicators for strain virulence selection.
Fungi ; Gene Expression ; Insect Proteins ; Peptide Hydrolases ; Virulence

OBJECTIVETo observe the apoptosis of neural stem cells (NSCs) at differential time points after the dissociation of neurospheres by Accutase or trypsin.

METHODSThe NSCs were isolated from striatum of human fetals that suffered abortion at 12-16 weeks of pregnancy. The 3(rd)-5(th) passages of NSCs were digested by Accutase or trypsin. Only vortexing was applied, and the triturating by Pasteur pipette was avoided to attenuate the injury to the cells during the dissociation. The single cells were then stained by Annexin V/propidium iodide and Hoechst 33342. The apoptosis rates 2 and 24 hours after passaging were evaluated.

RESULTSThe trypan blue staining confirmed that immediately after the dissociation,the viability of cells digested by trypsin was (83.10 ± 6.76)%, which was significantly lower than that digested by Accutase,which was (91.65 ± 4.43)% (P<0.05). The apoptosis of the NSCs digested by Accutase was higher than that digested by trypsin at both 2 and 24 hours after passaging (P<0.01). Four days after the passaging, both the new clone formation rate and diameter of new spheres after trypsin digestion were significantly higher than those after Accutase digestion (P<0.01).

CONCLUSIONSAlthough the viability of NSCs immediately after the disassociation by trypsin is lower than that digested by Accutase, the apoptosis of NSCs subsequently caused by trypsin is lower than that caused by Accutase. Trypan blue test immediately after the disassociation can not be used as an indicator in estimating the apoptosis of NSCs during the expanding.

PURPOSE: Ionic currents in smooth muscle have fundamental roles in initiation and maintenance of excitability and contractile status. So we studied and characterized potassium ion channels in smooth muscle cells of rabbit seminal vesicle. MATERIALS AND METHODS: Single smooth muscle cells of New Zealand White rabbit seminal vesicle were obtained using proteolytic enzymes (collagenase and papain). Using single cell and channel recording methods of patch clamp, potassium channels in smooth muscle of seminal vesicle were recorded. RESULTS: Potassium currents recorded whole-cell patch clamp method were divided into maxi-K channel dominant cells (n=11) and the cells with mixture of maxi-K and delayed rectifier K channels (n=7). Inside-out mode of patch clamp technique was used to characterize the maxi-K channel. The channel showed outward rectification and Ca-dependency. The single channel conductance of this channel estimated from slope conductance was 188 pS in physiological condition. These characters were typical properties of maxi-K channel. CONCLUSIONS: From these results it is suggested that maxi-K channel was predominantly distributed in rabbit seminal vesicle cell. The physiological roles of this channel in modulating seminal vesicle smooth muscle tone need further studies.
Humans ; Muscle, Smooth ; Myocytes, Smooth Muscle ; New Zealand ; Peptide Hydrolases ; Potassium ; Potassium Channels ; Seminal Vesicles

Atopic dermatitis (AD) is a multifactorial, chronic inflammatory skin disease in which genetic mutations and cutaneous hyperreactivity to environment stimuli play a role. The skin barrier is known to be damaged in patients with AD, both in acute eczematous lesions and in clinically unaffected skin. Skin barrier function can be impaired first by a genetic predisposition to produce increased levels of stratum corneum chymotryptic enzyme. This protease enzyme causes premature breakdown of corneodesmosomes, leading to damage of the epidermal barrier. The addition of environmental interactions can increase production of stratum corneum chymotryptic enzyme and impair epidermal barrier function. The epidermal barrier can also be damaged by exogenous proteases. One or more of these factors in combination might lead to a defective barriers, after then increasing the risk of allergen penetration and succeeding inflammatory reaction, thus contributing to exacerbations of this disease. The strong association between both genetic barrier defects and environmental factors to the barrier with AD suggests that epidermal barrier dysfunction is a primary event in the development of this disease. New concepts into the relation of the epidermal barrier function and its interaction with components of the innate and adaptive immune responses in patients with AD give rise to novel treatments.
Dermatitis, Atopic ; Genetic Predisposition to Disease ; Humans ; Peptide Hydrolases ; Skin ; Skin Diseases

Cereblon（CRBN) is a brain-associated protein with ionic protease activity, which interacts with DNA damage-binding protein-1 (DDB1), Cullin 4 (Cul4A or Cul4B), and regulator of Cullins 1 (RoC1) to form the functional E3 ubiquitin ligase complex(CRBN-CRL4) that performs proteolysis via the ubiquitin-proteasome pathway. And CRBN is a necessary target protein for the anti-myeloma effect of immunomodulators. The combination of lenalidomide and CRBN recruited a new substrate that binds to the CRBN-CRL4 complex, leading to increased ubiquitination and proteasome-dependent degradation, thus resulting in anti-myeloma activity. The substrates binding to this complex are IKZF1, IKZF3 proteins and GS, etc. The CRBN-dependent degradation of IKZF1 and IKZF3 after lenalidomide treatment is also the result of HO-mediated oxidative stress. In addition to ubiquitination, lenalidomide also mediates ubiquitin-independent pathways that prevent CRBN from binding to CD147-MCT1 in a competitive manner to regulate its antitumor activity. Lenalidomide can also play a role in multiple myeloma(MM) cells by modulating miRNA levels and CRBN binding to downstream protein AGO2 expression. Thus, there are many molecular mechanisms of lenalidomide anti-myeloma activity. This review summarizes the molecular mechanisms of CRBN in lenalidomide against myeloma activity in terms of ubiquitin-dependent and ubiquitin-independent pathways.
Cullin Proteins ; Humans ; Hydrogen Peroxide ; Multiple Myeloma ; Peptide Hydrolases ; Proteolysis ; Thalidomide ; Ubiquitination