• BMB Reports
• /
• v.41 no.2
• /
• pp.102-107
• /
• 2008

Extracellular proteases play an important role in a wide range of host physiological events, such as food digestion, extracellular matrix degradation, coagulation and immunity. Among the large extracellular protease family, serine proteases that contain a "paper clip"-like domain and are therefore referred to as CLIP-domain serine protease (clip-SP), have been found to be involved in unique biological processes, such as immunity and development. Despite the increasing amount of biochemical information available regarding the structure and function of clip-SPs, their in vivo physiological significance is not well known due to a lack of genetic studies. Recently, Drosophila has been shown to be a powerful genetic model system for the dissection of biological functions of the clip-SPs at the organism level. Here, the current knowledge regarding Drosophila clip-SPs has been summarized and future research directions to evaluate the role that clip-SPs play in Drosophila immunity are discussed.

• Journal of Microbiology
• /
• v.34 no.2
• /
• pp.198-205
• /
• 1996

Two proteases were partially characterized from culture filtrate of Mycobacterium, tuberculosis KIT110. Their molecular weights were approximately 200 and 180 kDa, respectively and they exhibited similar enzymatic characteristics. These enzymes were inhibited significantly by EDTA and to some extent by EGTA. Their activity was enhanced by $Ca^{2+}$ and $Mg^{2+}$ to some degree. However, $Cu^{2+}$ and $Ag^{2+}$ completely inhibited the enzyme activity at the concentration of 2.5 and 5 mM, respectively. The optimal pH was 7.0 and optimal temperature was around $40^{\circ}C$. These enzymes were rapidly inactivated at $80^{\circ}C$. Therefore, they were heat-labile, neutral metalloproteases. These enzymes exhibited antigenicity shown by their reacting with sera from the partients with pulmonary tuberculosis. These enzymes were able to degrade serum proteins including hemoglobin, bovine serum albumin, lysozyme and immunoglobulin G and structural matrix protein such as type I collagen. Therefore, these enzymes may be thought to contribute to tissue necrosis and pathogenesis during infection.

• Animal cells and systems
• /
• v.16 no.6
• /
• pp.431-437
• /
• 2012

Calpains are a class of proteins that belong to the calcium-dependent, non-lysosomal cysteine proteases. There are three major types of calpains expressed in the skeletal muscle, namely, ${\mu}$-calpain, m-calpain, and calpain 3, which show proteolytic activities. Skeletal muscle fibers possess all three calpains, and they are $Ca^{2+}$-dependent proteases. The functional role of calpains was found to be associated with apoptosis and myogenesis. However, calpain 3 is likely to be involved in sarcomeric remodeling. A defect in the expression of calpain 3 leads to limb-girdle muscular dystrophy type 2A. Calpain 3 is found in skeletal muscle fibers at the N2A line of the large elastic protein, titin. A substantial proportion of calpain 3 is activated 24 h following a single bout of eccentric exercise. In vitro studies indicated that calpain 3 can be activated 2-4 fold higher than normal resting cytoplasmic [$Ca^{2+}$]. Characterization of the calpain system in the developing muscle is essential to explain which calpain isoforms are present and whether both ${\mu}$-calpain and m-calpain exist in differentiating myoblasts. Information from such studies is needed to clarify the role of the calpain system in skeletal muscle growth. It has been demonstrated that the activation of ubiquitous calpains and calpain 3 in skeletal muscle is very well regulated in the presence of huge and rapid changes in intracellular [$Ca^{2+}$].

• The Korean Journal of Zoology
• /
• v.33 no.1
• /
• pp.119-125
• /
• 1990

A Potent inhibitor of trypsin and other various serine proteases including chymotrypsin, elastase, kallikrein, plasmin and subtilisin, has been purified to homogeneity from chick skeletal muscle by convendonal chromatographic procedures. The Inhibitor has an apparent molecular weight of 66, 000 dalton as determined by gel filtration. When the purified inhibitor was electrophoresed in the presence of sodium dodecyl sulfate, there appeared rwo protein bands having molecular weights of 66, 000 and 64, 000 dalton. The 64, 000 dalton protein seems to be the product of 66, 000 dalton protein by a lin'ited proteolysis during the purification procedure or in viuo. Thus, it seems to consist of a single polypeptide. The inhibitor appeared to be glycoprotein and have an isoelectric point of 7.4. It contains relatively large amount (8.33 mole%) of cysteine residues.

• Journal of Microbiology
• /
• v.37 no.1
• /
• pp.14-20
• /
• 1999

Two extracellular proteases, EP I and EP II, from cells of Oligotropha carboxydovorans (formerly Pseudomonas carboxydovorans) DSM 1227 grown in nutrient broth were purified to greater than 95% homogeneity in five steps using azocasein as a substrate. The final specific activities of EPs I and II were 214.9 and 667.4 units per mg of protein. The molecular weights of native EPs I and II were determined to be 23,000. Sodium dodecyl sulfate-gel electrophoresis revealed the two enzymes to be monomers. The enzymes were found to be serine-type proteases. The activity of EP I was stimulated by Ca2+, Mg2+, and Ba2+, but that of EP II was not. The enzymes were completely inhibited by Fe2+, Hg2+, Co2+, Zn2+, and Cd2+. EDTA and EGTA exhibited a strong inhibitory effect on EP I. The optimal pH for the two enzymes was pH 9.0. The optimal temperatures for EP I and II were 60 and 50$^{\circ}C$, respectively. The enzymes were stable under alkaline conditions. The thermal stability of EP I was higher than that of EP II. Cell-free extracts did not inhibit the purified enzymes. The enzymes were active on casein, azocasein, azocoll, and carbon monoxide dehydrogenase, but weakly active with bovine serum albumin.

• Korean Journal of Microbiology
• /
• v.33 no.2
• /
• pp.136-141
• /
• 1997

To elucidate whether there are bacteria excreting proteases in Korean traditional fermented food, soybean paste (Doen-Jang) or not, well growing bacteria with halos were isolated on the qkim milk agar media. The strains were identified as Bacillu, cereus JH-1, B. cereus SH-5, B. cereus SH-7 through various physiological and biochemical tests, VlTEK system, and MIDI system. The extracellular proteases of the strain JH-1, and SH-5, were optimal at pH 9, 40^{\circ}C.$, and the protease of strain SH-7 at pH 8 and 50^{\circ}C.$. Also hemolysis activities of the three strains were observed on the hlood agar media.

• Korean Journal of Food Science and Technology
• /
• v.43 no.6
• /
• pp.729-734
• /
• 2011

With the goal of transforming rice protein from an insoluble to a soluble form to increase the industrial utilization of rice wine meal (RWM), RWM was derivatized using commercial proteases and the RWM hydrolysates were characterized. Eight commercial proteases were used individually or in combination for hydrolysis of RWM. The degree of hydrolysis was assessed by determining the soluble protein in supernatant using the Lowry assay, protein in precipitates using a semimicro Kjeldahl procedure, and gravimetrically by the weight difference before and after hydrolysis. Protamex, Alcalase and Protease N proteases were most effective for hydrolysis of RWM. Although these assessment methodologies displayed some variation, they generally showed a similar pattern. When the aforementioned three proteases were simultaneously used to treat RWM, no significant difference was observed between the three assays (p<0.05) indicating an absence of enzymatic synergy.

• The Korean Journal of Food And Nutrition
• /
• v.16 no.4
• /
• pp.388-396
• /
• 2003

The freeze dried young antler residue was extracted by proteases and hydrochloric acid(HCl). The young antler was extracted by water at 50$^{\circ}C$ and the residue was reacted by proteases for 5 hours at 50$^{\circ}C$. The extraction rate of its residue was 32.8%(absorbance 3.61 at 280nm) of bacteria protease, 23.8%(absorbance 0.69) of papain, and 31.2% (absorbance 2.96) of pepsin. The young antler was extracted by boiling water and the residue was reacted by proteases for 5 hours at 50$^{\circ}C$. The extraction rate of its residue was 45.0%(absorbance 3.61) of bacteria protease, 30.4%(absorbance 0.33) of papain, and 51.2% (absorbance 2.77) of pepsin. The result of HPLC analysis reveals that in 50$^{\circ}C$ water extract and boiling water extract, all high molecular peak was reduced under MW 1,000 by proteases. The result from the extract of young antler residue reacted by HCl for 5 hours at 50$^{\circ}C$ shows that its extraction rate was 45% (absorbance 0.78) in concentration of 0.1N HCl, 61% (absorbance 1.82) in 0.2N, 81% (absorbance 2.29) in 0.4N, and 82.0% (absorbance 3.28) in 2.0N. The result of HPLC analysis also reveals that in the extract by 0.8N HCl, the peak of about MW 70,000 accounted for 78% in total. Protein content of the extract by 0.8N HCl was 8.2%, and content of amino acid was 81.6%, ash was 1.3%, and mineral contents were 0.1 % of Ca, 2.3% of P, 0.8 % of Mg, 3.4% of Na, 0.002% of F by dry base.

• Proceedings of the Korean Society for Applied Microbiology Conference
• /
• 2003.06a
• /
• pp.195-195
• /
• 2003

• The Zoological Society Korea : Newsletter
• /
• v.12 no.2
• /
• pp.111.1-111
• /
• 1995

No Abstract, See Full Text