• Journal of Life Science
• /
• v.25 no.1
• /
• pp.1-7
• /
• 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 Environmental Agriculture
• /
• v.22 no.3
• /
• pp.203-209
• /
• 2003

In the course of screening for antifungal agents, a bacterial strain, DM3 was isolated from a mud sample collected at Daechon in Chungnam province and identified as Bacillus licheniformis based on API 50CHB test. It has antifungal activity against 12 plant pathogenic fungi in paper disc assay. At the 95% lethal dose of gamma radiation ($^{60}Co$, 10 kGy, $D_{10}=2.32\;kGy$), the antifungal activity deficient mutant (mDM3) against Colletotrichum gloeosporioides was induced From 2-D electrophoresis analysis, serine hydroxymethyltransferase (45.0 kDa), hypothetical protein(40.7 kDa), NifU protein homolog(15.4 kDa), and resolvase(12.5 kDa) homologous proteins were detected only in B. licheniformis DM3. Lysozyme(18.1 kDa) and alkyl hydroperoxide reductase(15.6 kDa) homologous proteins were expressed uniquely in B. licheniformis mDM3. Further studies are needed to reveal that these proteins from B. licheniformis DM3 could be closely related to the antifungal activity against plant pathogenic fungi.

• Asian-Australasian Journal of Animal Sciences
• /
• v.21 no.6
• /
• pp.873-882
• /
• 2008

Eight ileally cannulated pigs (BW $35.9{\pm}0.9kg$) were randomly allotted according to a $4{\times}3$ Latin square design to determine the effects of cellulose, pectin and starch on standardized ileal digestibility (SID) and apparent total tract digestibility (ATTD) of crude protein (CP) and amino acids (AA) as well as on the bacterial AA contribution in feces. The pigs were fed the control diet (20.2% CP, % dry matter (DM)) or one of the three experimental diets in which 25% of the control diet was substituted by cellulose, starch or pectin. Due to this substitution, dietary CP levels were lower in the cellulose (15.5% CP, % DM), pectin (15.4% CP, % DM) and starch diet (15.2% CP, % DM). Following a 15-d adaptation period, feces were collected for 5 d and ileal digesta for a total of 24 h. Starch increased SID of CP, while cellulose and pectin had no significant effect on the digestibility of CP. Overall, starch supplementation resulted in higher (p<0.05) SID values of histidine, isoleucine, threonine, alanine, aspartic acid, cysteine, glycine and serine compared with cellulose, while pectin decreased (p<0.05) SID of valine and proline compared with the starch and control diet. Both cellulose and pectin reduced (p<0.05) the ATTD of CP and AA, while starch decreased (p<0.05) ATTD of phenylalanine, alanine, proline and serine compared with the control. With regard to bacterial AA composition of the fecal mixed bacterial mass (MBM), cellulose supplementation increased (p<0.05) its content of N and almost all AA, except for valine, while pectin caused higher contents of arginine, histidine and proline compared with the control (p<0.05). The bacterial contribution of arginine in feces was higher (p<0.05) in the cellulose treatment, while pectin reduced (p<0.05) the bacterial contribution of leucine, alanine, glutamic acid and proline in feces compared with the control. In conclusion, the effects of cellulose, starch and pectin on SID were rather small. Bacterial activity in the large intestine can only explain the reduced ATTD values for arginine in the cellulose treatment, but not for the other AA in the cellulose and pectin treatments, suggesting higher endogenous losses of these AA in the large intestine.

• 한국해양학회지
• /
• v.30 no.4
• /
• pp.341-354
• /
• 1995

Fluorescence characteristic and amino acids composition of organic matter were determined from extracted seawater samples at eight stations in the East Sea of Korea. Organic compounds have been extracted onto C-18 Sep-Pak cartridges. Three dimensional excitation/emission fluorescence contouring of extracts showed two markedly distinct characterized fluoroscopies representing protein-like biomacromolecule and humic-like geomacromolecule. Protein-like biomacromolecule showing fluorescence maxima at 280 nm/330 nm (excitation/emission) were abundant in the surface mixed layer and then apparently decreased below the thermocline at most stations. It suggests that source of biomacromolecule is comely related with vigorous biological synthetic activity in the surface layer and bacteria decompose its biologically labile components near the thermocline and in the deeper layer. On the other hand, humiliate geomacromolecule showing fluorescence maxima at 330 nm/430 nm (excitation/emission) were low in the surface mixed layer implying photochemical oxidation and then increased below the thermocline at most stations. It suggests that geomacromolecule might be transformed by condensation of bio-refractoryorganic fraction after decomposition of biomacromolecule and particulate organic carbon derived from the surface mixed layer. HPLC measurements of amino acids showed similar composition between seawater and extracted organic macromolecule after hydrolysis. Glycine, serine and alanine were predominant, accounting for more than 50% of total amino acids. Dissolved free amino acids of seawater were more abundant in the surface layer(0.7∼1.8 uM) than the deeper layer (0.2∼0.4 uM). D/L racemic ratio of alanine of extracted organic matter showed lower value in the surface layer than the deeper layer. It suggests that biomacromolecule predominant in the surface layer is relatively young, rapidly recycling and biologically labile.

• Proceedings of the Korean Society of Applied Pharmacology
• /
• 2002.07a
• /
• pp.236-236
• /
• 2002

Pyruvate dehydrogenase phosphatase (PDP) is a mitochondrial protein serine/threonine phosphatase that catalyzes the dephosphorylation and concomitant reactivation of the pyruvate dehydrogenase componant of the pyruvate dehydrogenase complex (PDC). PDP consists of a Mg$\^$+2/ -dependent and Ca$\^$+2)-stimulated catalytic subunit (PDPc) of Mr 52,600 and a FAD-containing regulatory subunit (PDPr) of Mr 95.600. Catalytic subunit of pyruvate dehydrogenase phosphatase (PDPc) has been suggested to have three major functional domains such as dihydrolipoamide acetyltransferase(E$_2$)-binding domain, regulatory subunit of PDP(PDPr)-binding domain, and calcium-binding domain. In order to identify functional domains, recombinant catalytic subunit of pyruvate dehydrogenase phosphatase (rPDPc) was expressed in E. coli JM101 and purified to near homogeneity using the unique property of PDPc: PDPm binds to the inner lipoyl domain (L$_2$) of E$_2$ of pyruvate dehydrogenase complex (PDC) in the presence of Ca$\^$+2/, not under EGTA. PDPc was limited-proteolysed by trypsin, chymotrypsin, Arg-C, and elastase at pH7.0 and 30$^{\circ}C$ and N-terminal analysis of the fragment was done. Chymotrypsin, trypsin, and elastase made two major framents: N-terminal large fragment, approx. 50kD and C-terminal small fragment, approx. 0 kDa. Arg-C made three major fragments: N-terminal fragment, approx. 35 kD, and central fragment, approx. 15 kD, and C-terminal fragment, approx. 10 kD. This study strongly suggest that PDPc consists of three major functional domains. However, further study should be necessary to identify the functional role.

• Korean Journal of Veterinary Research
• /
• v.43 no.3
• /
• pp.439-448
• /
• 2003

The VP2 gene of infectious bursal disease virus (IBDV) Chinju which was previously detected in Chinju, Korea was cloned and sequenced to establish the information for the development of genetically engineered vaccines and diagnostic reagents against IBDV. The nucleotide sequence of the entire Chinju VP2 gene consisted of 1,356 bases long encoding 452 amino acids in a single open reading frame (ORF). It consisted of 368 adenine (27.1%), 363 cytosine (26.8%), 339 guanine (25.0%) and 286 thymine (21.1%) residues. The predicted $M_r$ of the Chinju VP2 protein was 48 kDa, and the protein contained 13 phosphorylation sites by protein kinase C, casein kinase II or tyrosine kinase, whereas 3 asparagine-linked glycosylation sites were recognized. The nucleotide sequence of Chinju VP2 ORF had a very close phylogenetic relationship with 98-99% homology to that of the very virulent IBDVs (vvIBDVs) HK46, OKYM, D6948, UK661, UPM97/61 and BD3/99. Also, the Chinju VP2 protein revealed a very close phylogenetic relationship with 99-100% homology to that of these vvIBDVs. The Chinju VP2 protein had 100% amino acid identity in the variable region of residues 206-360 with that of the D6948, HK46, OKYM and UK661, as well as 100% identity in two hypervariable regions of residues 212-224 and 314-324 with those of the D6948, HK46, OKYM, UK661, UPM97/61 and BD3/99. The amino acid sequence of the chinju VP2 protein contained a serine-rich heptapeptide of SWSASGS as in these vvIBDVs.

• Journal of the Korean Chemical Society
• /
• v.30 no.2
• /
• pp.216-223
• /
• 1986

Separation of optical isomers of DNS derivatized amino acids by a reversed-phase high-performance liquid chromatography has been studied by adding a complex of an optically active amino acid (L-arginine) with the metal ion (Cu(II), Zn(II), Cd(II), Ni(II)) to the mobile phase. The separations are affected by the concentrations of acetonitrile, chelate and buffer. They are also affected by the pH and the kinds of metal and buffer. A separation mechanism, which is based on steric effect of the ligand exchange reaction for the formation of ternary complexes by the D,L-DNS-amino acids and the chiral additive associated with the stationary phase, is proposed to interpret the elution behaviors of D, L-dansyl-amino acids.

• Korean Journal of Environmental Biology
• /
• v.22 no.1
• /
• pp.148-152
• /
• 2004

This study was carried out to investigate the callus formation and to determine the composition and contents of the Hee amino acids in seedlings and callus of Rhodiola sachalinensis. The callus formation from the part of seedling-roots was the most effective on MS (Murashige and Skoog) media supplemented with 2.0 mg $L^{-1}$2,, 4-D included in 1.0 mg $L^{-1}$ kinetin than the other experimental plots which prepared with the different concentration of the various growth regulators. Free amino acids extracted from the explants and the callus were a total of 25∼26 kinds. Especially, the basic amino acids such as arginine, lysine and histidine released in callus were found relatively in a large quantity. These results suggest that the cultured callus of Rhodiola sachatinensis could induce for the mass production of the other useful ingredients.

• Korean Journal of Veterinary Pathology
• /
• v.2 no.2
• /
• pp.73-84
• /
• 1998

This study was carried out to investigate on the serum chemistry and the DNA ploidy changes in carcinogenesis of the rat liver and kidney. Sixty male Sprague-Dawley rats were divided into two groups. Group I was non-treated control. Group II was given initiators (2,2'-dihydroxy- di-N-propylnitrosamine, 0.1% in drinking water(d.w.) for 1 week and N-ethyl-N-hydroxy-ethylnitrosamine; 0.15% in d.w. for 1 week) and promoters (3'methyl-cholanthrene; 3'MC, l0mg/kg, intraperitoneally(i.p.) twice a week and DL-serine; 0.05% in d.w. for 5 weeks, from 3 to 8 weeks). All examinations were performed at 12 and 20 weeks RBC, HGBCp＜0.05) and PCVCp＜0.01) significantly decreased in Group II at 20 weeks. Activities of ALT, AST(p＜0.05) and GGT(p＜0.01) were significantly increased in Group II at 20 weeks. Flow cytometric analysis showed hepatocyte nuclei from normal livers were predominantly tetraploid(66~67%) and then diploid(28~30%). Most of hepatocyte nuclei from carcinogen-treated rats were diploid (52~68%) and less were tetraploid(28~42%). Neoplastic liver nodules and hepatocellular carcinoma contained almost exclusively diploid nuclei. Renal cell nuclei from normal kidney were predominantly diploid(88~93%), those from carcinogen-treated rats had an abnormal DNA-content peak(aneuploidy, 6-7%), near the tetraploidy area. These results suggest that diploidy may be an effective screening marker of the liver carcinogenesis. Aneuploidy may be an useful marker in assessment of the experimental renal carcinogenesis.

• Reproductive and Developmental Biology
• /
• v.33 no.4
• /
• pp.243-248
• /
• 2009

To examine the differential expression of proteins during the cycling (70~80% confluences) and G0/G1 (full confluences) phases in porcine fetal fibroblast cells, we used a global proteomics approach by 2-D gel electrophoresis (2-DE) and MALDI-TOF-MS. Cycling cell were harvested at approximately 70% to 80% confluent state while cells in G0/G1 phase were recovered after maintenance of a confluent state for 48 hr. Cellular proteins with isoelectric points ranging between 3.0~10.0, were analyzed by 2-DE with 2 replicates of each sample. A total of approximately 700 spots were detected by 2.D gels stained with Coomassie brilliant blue. On comparing the cell samples obtained from the cycling and G0/G1 phases, a total of 13 spots were identified as differentially expressed proteins, of which 8 spots were up-regulated in the cycling cell and 5 were up-regulated in the G0/G1 phase. Differentially expressed proteins included K3 keratin, similar to serine protease 23 precursor, protein disulfide-isomerase A3, microsomal protease ER-60, alpha-actinin-2, and heat-shock protein 90 beta. The identified proteins were grouped on the basis of their basic functions such as molecular binding, catabolic, cell growth, and transcription regulatory proteins. Our results show expression profiles of key proteins in porcine fetal fibroblast cells during different cell cycle status.