• Biomedical Science Letters
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• v.13 no.4
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• pp.263-272
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• 2007

Nebulin is a giant actin binding protein (600-900 kDa) which is specific to skeletal muscle. This protein is known to regulate thin filaments length in sarcomere as a molecular template. The C-terminus of nebulin is located in the Z-disc of muscle sarcomere and is bound to other proteins such like myopalladin, titin, archvillin, and desmin. The N-terminus of nebulin binds to tropomodulin at the pointed ends of the thin filaments. In recent research, nebulin not only found in brain but also expressed in heart, stomach, and liver. So, the roles of nebulin in non-muscle tissue have been studied. However, lack of information or studies on nebulin binding proteins and nebulin function in brain are available so far. Therefore, the current study have investigated a novel binding partner of Nebulin C-terminus by using yeast two-hybrid screening with human brain cDNA library. Nebulin C-terminus, containing simple repeats, serine rich and SH3 domain, interacts with osteonectin C-terminal region. The specific interaction of nebulin and osteonectin were confirmed in vitro by using GST pull-down assay and reconfirmed in vivo by using transfected COS-7 cells with EGFP-tagged nebulin and DsRed-tagged osteonectin. Consequently, this study identified SH3 domain in nebulin C-terminus specifically binds to extracellular Ca-binding (EeC domain in osteonectin. Also, nebulin C-terminus fusion protein colocalized with osteonectin EC domain fusion protein in transfected COS-7 cells. The current study found the interaction between nebulin and osteonectin in human brain for the first time and suggested the nebulin in brain may be associated with osteonectin, as a regulator of cell cycle progression and mitosis.

• Journal of the Korean Society of Food Science and Nutrition
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• v.13 no.4
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• pp.406-412
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• 1984

Total lipids (TL) from Korean pinenut (Pinuskoraiensis S & Z) were extracted, purified and fractionated into three lipid classes (neutral lipid: NL, glycolipid; GL, phospholipid; PL). Lipid contents(constituent components) and fatty acid composition of three lipid classes were determined by thin layer chromatography and gas chromatography. TL ranged from 69.0% to 69.8% in fresh pinenut and consisted of 95.9% to 96.7% NL, 3.2% to2.5% GL and 0.9% to 0.8% PL. In the NL, triglycerides were predominant (80.8%) with the smaller amounts of sterol, diglycerides, free fatty acids, sterol esters and hydrocarbons. Monogalactosyl diglycerides and esterified steryl glycosides (23.5%) were the major components of GL, but cerebrosides, steryl glycosides and digalactosyl diglycerides were also found as minor components. Of the PL, phosphatidyl choline (40.2%) and phosphatidyl ethanolamine (19.4%) were the major components, comprising over 60% of this class. Phosphatidyl inositol, lysophosphatidyl choline were also present in the PL. The major fatty acids in the NL were linoleic acid (48.6%), oleic acid (28.8%) and arachidic acid(14.4%), The fatty acid composition in the GL was similar to the pattern in the NL, but PL contained a higher percentage of palmitic acid (17.7%) and stearic acid (6.0%) than other lipid classes.

• Childhood Kidney Diseases
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• v.18 no.2
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• pp.98-105
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• 2014

Purpose: Nafamostat mesilate (NM), a synthetic serine protease inhibitor, has been investigated as an anticoagulant for adult patients with a high risk of bleeding, who need chronic renal replacement therapy (CRRT). However, little is known about the use of NM as an anticoagulant in pediatric CRRT. The aim of this study was to evaluate the ideal dosage, efficacy, and safety of NM in pediatric CRRT. Methods: We conducted a retrospective study of 40 pediatric patients who had undergone at least 24 h of venovenous CRRTs between January 2011 and October 2013. We divided the patients according to risk of bleeding. Those at high risk received no anticoagulation (group 1) or NM as an anticoagulant (group 2), while those at low risk received heparin (group 3). Results: Forty patients (25 male and 15 female; mean age, $8.2{\pm}6.6$ years) were enrolled. The mean duration of CRRT was 13.0 days, and the survival rate was 57.5%. The mean hemofilter lifespan was 39.3 h in group 1 and 11.3 h in group 3. In group 2, hemofilter lifespan was extended from 7.5 h to 27.4 h after the use of NM (P =0.001). The mean hemofilter lifespan with NM was greater than with heparin (P =0.018). No patient experienced a major bleeding event during treatment with NM. Conclusion: NM may be a good alternative anticoagulant in pediatric patients with a high risk of bleeding requiring CRRT, and is not associated with bleeding complications.

• Korean Journal of Weed Science
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• v.13 no.2
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• pp.81-88
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• 1993

For searching the photosynthetic electron transport(PET) inhibitors, bio-rational screening system using thylakoid membranes extracted from wild and mutant cyanobacteria(Anacystis nidulans $R_2$) was developed. Generally, thylakoid membrane was more sensitive to the tested herbicides than the chloroplast from spinach in the Hill reaction. Higher resistant characteristics appeared in mutant D-5, Di-22 to diuron and mutant G-264 to atrazine as compared to wild type. To test the reaction of thylakoid membrane to herbicides, diuron and atrazine were applied simultaneously. Diuron and atrazine competed each other for binding with substituted amino acids, while diuron and dinoseb were non-competitive, and inhibiting activity was increased. Conclusively, bio-rational screening system using cyanobacteria was proved to be fast and efficient screening method for the development of PET inhibitors.

• Journal of Mushroom
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• v.18 no.1
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• pp.63-71
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• 2020

The present study aimed to investigate the biological activities and nutritional contents of extracts from three types of edible mushrooms (Pleurotus ostreatus, P. eryngii, and Hericium erinaceus) following different roasting treatments. Roasting by a household multi-oven (HR60) decreased the total polyphenol contents, as well as DPPH radical scavenging and nitrite scavenging activities compared with the fresh samples, and no significant difference was observed between the different roasting times. As a result of rotary drum roasting (CR) treatment, we could observe a decrease in the DPPH radical scavenging activity in all roasted mushroom samples compared with the fresh samples, whereas nitrite scavenging activity increased in roasted P. eryngii and H. erinaceus. Additionally, total polyphenol contents increased significantly in roasted H. erinaceus. In each roasted sample dried by HR60 and CR, the amount of palatable-taste amino acids (e.g., glutamic acid) and sweet-taste amino acids, including glycine, serine, alanine, and threonine, increased in P. ostreatus, whereas the amount of bitter amino acids, including leucine, isoleucine, histidine, valine, and methionine, decreased compared with the fresh samples. In P. eryngii and H. erinaceus samples, we also observed an increase in the amount of palatable and sweet-taste amino acids upon the roasting treatments; however, the amount of bitter amino acids increased simultaneously following the roasting treatments. These results suggest that CR treatment is more effective in increasing the biological activities and nutritional contents, such as palatable and sweet-taste amino acids, compared with HR60 treatment.

• Journal of Life Science
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• v.25 no.1
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• pp.1-7
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• 2015

Cyclin D is a member of the cyclin protein family, which plays a critical role as a core member of the mammalian cell cycle machinery. D-type cyclins (D1, D2, and D3) bind to and activate the cyclin-dependent kinases 4 and 6, which can then phosphorylate the retinoblastoma tumor suppressor gene products. This phosphorylation in turn leads to release or derepression of E2F transcription factors that promote progression from the G1 to S phase of the cell cycle. Among the D-type cyclins, cyclin D3 encoded by the CCND3 gene is one of the least well studied. In the present study, we have investigated the biochemistry of the proteolytic mechanism that leads to loss of cyclin D3 protein. Treatment of human prostate carcinoma PC-3-M cells with lovastatin and actinomycin D resulted in a loss of cyclin D3 protein that was completely reversible by the peptide aldehyde calpain inhibitor, LLnL. Additionally, using inhibitors for various proteolytic systems, we show that degradation of cyclin D3 protein involves the $Ca^{2+}$-activated neutral protease calpain. Moreover, the half-life of cyclin D3 protein half-life increased by at least 10-fold in PC-3M cells in response to the calpain inhibitor. We have also demonstrated that the transient expression of the calpain inhibitor calpastatin increased cyclin D3 protein in serum-starved NIH 3T3 cells. These data suggested that the function of cyclin D3 is regulated by $Ca^{2+}$-dependent protease calpain.

• Korean Journal of Food Science and Technology
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• v.25 no.5
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• pp.421-425
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• 1993

Hololeion maximowiczii root was analyzed for proximate composition, free sugar, fatty acids of crude lipid, amino acids and minerals after freeze dried (FD), hot air dried (HAD) and hot air dried after blanched (HADB), respectively. Proximate composition was little different among drying methods. Free sugar was abundant in order of sucrose $(28{\sim}29mg/g\;dw)$, fructose $(0.8{\sim}3mg/g\;dw)$, glucose and xylose (trace). Fructose was decreased in HADB. In fatty acid composition of crude lipids, linoleic acid was highest $(55{\sim}59%)$ and then palmitic and linolenic acid in abundance. These three fatty acids made $88{\sim}90%$ of total lipid, Low molecular fatty acids showed decreasing tendancy in HAD and HADB. Amino acid composition of hydrolyzed sample showed abundancy in order of arginine $(34{\sim}42mg/g)$, lysine, glutamic acid, glycine, aspartic acid, valine and histidine, while that of free amino acids was arginine, histidine, lysine, aspartic acid, serine, glutamic acid, isoleucine. Glycine and glutamic acid in hydrolyzed sample showed slight decreasing tendency in HADB. Mineral content was abundance in order of Ca $(1927{\sim}2379\;ppm),\;K\;(1159{\sim}1259ppm)$ and $Mg(760{\sim}777\;ppm)$. No difference was found in drying methods.

• Applied Biological Chemistry
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• v.12
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• pp.43-51
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• 1969

The purpose of this study was to determine protein amino acid contents of some Korean foods by gas-liquid chromatography, and to evaluate this technique as a procedure for the quantitative determination of amino acids in foods. The crude protein content of foods was also estimated from the nitrogen content. 1. Nitrogen content of each food sample was determined previously to adjust the amount of sample for GLC analysis 2. In the analysis of 17 known amino acids, a linear relationship was found between the weight of 13 amino acids of 17 amino acids, the internal standard as well as the injection volume of a mixture and the detector responses for the derivatives of the amino acids. No response for arginine, cystein, histidine, and tyrosine was observed. 3. The relative molar response (RMR) values for the 13 amino acids of standard solution relative to glutamic acid as '1.00' were obtained under normal operating conditions with a hydrogen flame ionization detector. 4. The recovery of amino acids from their mixtures with natural food materials was carried out. The recoveries were essentially quantitative except threonine and serine. An overall mean recovery of 11 amino acids was $101.4{\pm}8.4$ per cent before hydrolysis and $98.1{\pm}8.7$ per cent after hydrolysis of samples. 5. The comparative analysis of the acid hydrolysates of two food samples by gas-liquid and ion-exchange chromatographic analysis were carried out. In white-bait pemmican, only threonine and asparagine amounts by GLC analysis had similar values to those obtained by ion-exchange chromatography. The other seven amino acids gave higher values as measured by GLC than by ion-exchange. With the food sample, soybean, alanine, valine, asparagine, and glutamic acid were in good agreement in two analysis, while leucine, proline, threonine, phenylalanine, and lysine were found in slightly higher concentrations in the GLC analysis. 6. Grant variations of amino acid content were found among samples analyzed. The amino acid contents of each sample were compared with the values found in the literature.

• Applied Biological Chemistry
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• v.13 no.1
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• pp.65-72
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• 1970

Two peptides (Im pr-M, Im ch-M) derived from Im ${\lambda}-type$ of Bence Jones Protein and one peptide (Ikch-M) from Ik were separated and purified using the Dowex $50{\times}2$ column $(1{\times}20\;cm)$ and Dowex $1{\times}2(0.9{\times}50\;cm)$. The buffer solution was composed of 1% pyridine and IM formic acid in Dowex $1{\times}2$ column. The blocked N-terminal was examined with ninhydrin reaction before and after alkaline hydrolysis, which was fractionated by Dowex $1{\times}2$ column. Pyrro-glutamic acid in N-terminal residue was identified by comparing with the authentic pyrro-glutamic acid through a high voltage electrophoresis (pH 3.5, 3000 V.) after the peptide Im pr-M (PCA. Ser) was cleavaged at the position of serine with cone. (12 N) HCl and the pyrro-glutamic acid was converted to glutamic acid by treating it with N-NaOH for 116 hours at $27^{\circ}C$. The substractive method was applied to find out the sequence of peptides and carboxypeptidase A was employed to release C-terminal residue from the peptide. In present study PCA. Ser in Im Pr-M was isolated from the pronase digested ${\lambda}$-type Bence Jones protein. The yield of the Im Pr-M was 79.6 percent of its theoretical value, based on the molecular weight of Bence Jones Protein. Im ch-M (PCA. Ser Val. Leu) was isolated from the chymotrypsin digested ${\lambda}$-type Bence Jones Protein. The yield of the Im ch-M was 72.2 percent. based on the molecular weight of Bence Jones Protein. Ik ch-M (PCA. Ser. Ala. Leu) was isolated from the chymotrypsin digested ${\lambda}$-type Bence Jones Protein and its yield was 42% based on the molecular weight of Bence Jones Protein.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2002.07a
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• pp.113-113
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• 2002

Phylogenetically conserved Bcl-2 family proteins play a pivotal role in the regulation of apoptosis from virus to human. Members of the Bcl-2 family consist of antiapoptotic proteins such as Bcl-2, Bcl-xL, and Bcl-w, and proapoptotic proteins such as BAD, Bax, BOD, and Bok. It has been proposed that anti- and proapoptotic Bcl-2 proteins regulate cell death by binding to each other and forming heterodimers. A delicate balance between anti- and proapoptotic Bcl-2 family members exists in each cell and the relative concentration of these two groups of proteins determines whether the cell survives or undergoes apoptosis. Mcl-1 (Myeloid cell :leukemia-1) is a member of the Bcl-2 family proteins and was originally cloned as a differentiation-induced early gene that was activated in the human myeloblastic leukemia cell line, ML-1 . Mcl-1 is expressed in a wide variety of tissues and cells including neoplastic ones. We recently identified a short splicing variant of Mcl-1 short (Mcl-IS) and designated the known Mcl-1 as Mcl-1 long (Mcl-lL). Mcl-lL protein exhibits antiapoptotic activity and possesses the BH (Bcl-2 homology) 1, BH2, BH3, and transmembrane (TM) domains found in related Bcl-2 proteins. In contrast, Mcl-1 S is a BH3 domain-only proapoptotic protein that heterodimerizes with Mcl-lL. Although both Mc1-lL and Mcl-lS proteins contain BH domains fecund in other Bcl-2 family proteins, they are distinguished by their unusually long N-terminal sequences containing PEST (proline, glutamic acid, serine, and threonine) motifs, four pairs of arginine residues, and alanine- and glycine-rich regions. In addition, the expression pattern of Mcl-1 protein is different from that of Bcl-2 suggesting a unique role (or Mcl-1 in apoptosis regulation. Tankyrasel (TRF1-interacting, ankyrin-related ADP-related polymerasel) was originally isolated based on its binding to TRF 1 (telomeric repeat binding factor-1) and contains the sterile alpha motif (SAM) module, 24 ankyrin (ANK) repeats, and the catalytic domain of poly(adenosine diphosphate-ribose) polymerase (PARP). Previous studies showed that tankyrasel promotes telomere elongation in human cells presumably by inhibiting TRFI though its poly(ADP-ribosyl)action by tankyrasel . In addition, tankyrasel poly(ADP-ribosyl)ates Insulin-responsive amino peptidase (IRAP), a resident protein of GLUT4 vesicles, and insulin stimulates the PARP activity of tankyrase1 through its phosphorylation by mitogen-activated protein kinase (MAPK). ADP-ribosylation is a posttranslational modification that usually results in a loss of protein activity presumably by enhancing protein turnover. However, little information is available regarding the physiological function(s) of tankyrase1 other than as a PARP enzyme. In the present study, we found tankyrasel as a specific-binding protein of Mcl-1 Overexpression of tankyrasel led to the inhibition of both the apoptotic activity of Mel-lS and the survival action of Mcl-lL in mammalian cells. Unlike other known tankyrasel-interacting proteins, tankyrasel did not poly(ADP-ribosyl)ate either of the Mcl-1 proteins despite its ability to decrease Mcl-1 proteins expression following coexpression. Therefore, this study provides a novel mechanism to regulate Mcl-1-modulated apoptosis in which tankyrasel downregulates the expression of Mcl-1 proteins without the involvement of its ADP-ribosylation activity.