• BMB Reports
• /
• v.41 no.12
• /
• pp.881-885
• /
• 2008

Protein phosphatase 1 consists of a secondary structure arrangement, conserved in the serine/threonine protein phosphatase gene family, flanked by nonconserved N-terminal and C-terminal domains. The deletion mutant of PP1 with the 8 nonconserved N-terminal residues removed was designated PP1-(9-330). PP1-(9-330) had a higher activity and affinity than PP1 when assayed against four different substrates, and it also demonstrated a 6-fold higher sensitivity to the inhibitor okadaic acid. This suggested that the N-terminal domain suppresed the activity of PP1 and interfered with its inhibition by okadaic acid. The ANS fluorescence intensity of PP1-(9-330) was greater than that of PP1, which implies that the hydrophobic groove running from active site in the truncated PP1 was more hydrophobic than in PP1. Our findings provide evidence that the nonconserved N-terminus of PP1 functions as an important regulatory domain that influences the active site and its pertinent properties.

• 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.

• Proceedings of the Korean Society of Food Hygiene and Safety Conference
• /
• 2002.05a
• /
• pp.215-215
• /
• 2002

Catalytic subunit of pyruvate dehydrogenase phosphatase (PDPc) has been suggested to have three major funational domains such as dihydrplipoamide adetyltransferase(E2)-binding domain, regulatory subunit of PDP(PDP)r-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: PDPc binds to the inner lipoyl domain (L2) of E2 of ppyruvate dehydrogenase complex (PDC) in the presence of Ca+2, not under EGTA. PDPc was limited-proteolysed by typsin, chymotypsin, Arg-C, and elastase at pH 7.0 and 30C and N-terminal analysis of the fragments was done. Chymotrypsin, trypsin, and elastase made two major fragments: N-terminal large fragment, approx. 50kD and C-terminal small fragment, approx.10 kDa. Arg-C made three major fragments: N-terminal fragment, approx. 35kD, 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.

• Journal of Periodontal and Implant Science
• /
• v.39 no.2
• /
• pp.149-156
• /
• 2009

Purpose: Deproteinized bovine bone substitutes are commonly used in dental regenerative surgery for treatment of alveolar defects. In this study, three different bovine bone minerals - OCS-B (NIBEC, Seoul, Korea), Bio-Oss (Geistlich - Pharma, Switzerland), Osteograft/N - 300 (OGN, Dentsply Friadent Ceramed. TN, USA) - were investigated to analyze the basic characteristics of commercially available bone substitutes. Methods: Their physicochemical properties were evaluated by scanning electron microscopy, energy dispersive X-ray spectrometer (EDS), surface area analysis, and Kjeldahl protein analysis. Cell proliferation and alkaline phosphatase (ALP) activity of human osteosarcoma cells on different bovine bone minerals were evaluated. Results: Three kinds of bone substitutes displayed different surface properties. Ca/P ratio of OCS - B shown to be lower than other two bovine bone minerals in EDS analysis. Bio-Oss had wider surface area and lower amount of residual protein than OCS - B and OGN. In addition Bio - Oss was proved to have lower cell proliferation and ALP activity due to lots of residual micro particles, compared with OCS - B and OGN. Conclusions: Based on the results of this study, three bovine bone minerals that produced by similar methods appear to have different property and characteristics. It is suggested that detailed studies and quality management is needed in operations for dental use and its biological effects on new bone formation.

• Journal of Periodontal and Implant Science
• /
• v.27 no.2
• /
• pp.291-303
• /
• 1997

Periodontal ligament cells may have a role in the regulation of hard and soft periodontal tissues, but their specific function has not yet to be determined. To evaluate further their role in periodontal regeneration, they were examined for osteoblast-like behavior. Periodontal ligament cells and gingival fibroblasts were primarily cultured from extracted premolar with non-periodontal diseases. Cells were cultured with DMEM at $37^{\circ}C$, 5% $CO_2$, 100% humidity incubator, and as a measure of cell characterization, it was examined that the morphology, alkaline phosphatase activity, collagen synthesis, and immunocytochemistry for osteonectin, osteocalcin, and collagen type I. Healthy periodontal ligament cells has more osteoblastic-like cell property in alkaline phosphatase activity. and collagen synthesis than gingival fibroblast. Immunocytochemistry localization explained that calcitonin were expressed in periodontal ligament cells only, and osteonectin and type I collagen were produced in both cells simultaneously. This results indicate that the growth characteristics of periodontal ligament cells and gingival fibroblasts exhibit some differences in proliferative rates and biochemical synthesis. The differences may help to calrify the role such cells play in the regenearation of periodontal tissues.

• Advances in Traditional Medicine
• /
• v.3 no.3
• /
• pp.133-140
• /
• 2003

Petroleum ether, chloroform and ethanol extracts of the seeds of Melia azedarach Linn. administered orally to male rats at the dose level of 25 mg/100 g body weight for 48 days showed antispermatogenic activity, as the number of spermatocytes, spermatids and spermatozoa was decreased. The total cholesterol content was increased while protein and glycogen contents were decreased. The acid phosphatase content was also decreased while that of alkaline phosphatase increased. At the same time the weight of caput and cauda epididymis, prostate gland, seminal vesicle and Levator Ani muscle was decreased indicating its antiandrogenic property. Of the three extracts, the petroleum ether extract was more potent in its antispermatogenic and antiandrogenic activities and did not produce any sings of toxicity upto a dose of 25 mg/100 g body weight oral administration. After subjecting it to preliminary phytochemical screening the petroleum ether extract showed positive tests for steroids and saponins.

• Bulletin of the Korean Chemical Society
• /
• v.28 no.7
• /
• pp.1171-1174
• /
• 2007

We have developed an electrochemical immunosensor that combines the electrocatalytic property of carbon nanotube and the low background current of indium tin oxide (ITO) electrode. A partial monolayer of carboxylated single-walled carbon nanotube (CCNT) is covalently formed on an ITO electrode modified with amine-terminated phosphonic acid. Nonspecifically adsorbed avidin on the hydrophobic sidewalls of CCNT is used to immobilize biotinylated antibody and to reduce the nonspecific binding to CCNT. The biotinylated antimouse IgG bound on avidin and the antimouse IgG conjugated with alkaline phosphatase (ALP) sandwiches a target mouse IgG. ALP catalyzes the conversion of p-aminophenyl phosphate monohydrate into p-aminophenol, which is electrocatalytically oxidized to p-quinone imine on CCNT surface. Moderate electrocatalytic electrode obtained with the combination of CCNT and ITO allows low detection limit (0.1 ng/ mL).

• Journal of Periodontal and Implant Science
• /
• v.38 no.4
• /
• pp.595-602
• /
• 2008

Purpose: Ti-6Al-4V alloy is widely used as an implant material because of its good biocompatibility and good mechanical property compared with commercial pure titanium. Otherwise, toxicity of aluminum and vanadium in vivo has been reported. Ti-8Ta-3Nb alloy is recently developed in the R&D Center for Ti and Special Alloys and it was reported that this alloy has high mechanical strength, no cytotoxicity and similar biocompatibility to commercial pure titanium, but many studies are needed for its clinical use. In these experiment, we carried out different surface treatment on each Ti-8Ta-3Nb alloy disks, then cultured cell on it and assessed biological response. Materials and Methods: cpTi, Ti-6Al-4V, Ti-8Ta-3Nb alloy disks were prepared and carried out sandblasting and acid etching (SLA) or alkali-heat treatment (AH) on the Ti-8Ta-3Nb alloy disks. We cultured primary rat calvarial cells on each surface and assessed early cell attachment and proliferation by scanning electron microscopy, cell proliferation, alkaline phosphatase activity. Result: The rates of cell proliferation on the cpTi, Ti-8Ta-3Nb AH disks were higher than others (p<0.05) and alkaline phosphatase activity was significantly enhanced on the Ti-STa-8Nb AH disks(p<0.05). Conclusion: Most favorable cell response was shown on the Ti-8Ta-3Nb AH surfaces. It is supposed that alkali-heat treatment of the Ti-8Ta-3Nb alloy could be induced earlier bone healing and osseointegration than smooth surface.

• Journal of Ginseng Research
• /
• v.37 no.4
• /
• pp.435-441
• /
• 2013

This study investigated the effects of Korean Red Ginseng (KRG) on radiation-induced bone loss in C3H/HeN mice. C3H/HeN mice were divided into sham and irradiation (3 Gy, gamma-ray) groups. The irradiated mice were treated for 12 wk with vehicle, KRG (per os, p.o.) or KRG (intraperitoneal). Serum alkaline phosphatase (ALP), tartrate-resistant acid phosphatase, estradiol level, and biomechanical properties were measured. Tibiae were analyzed using micro-computed tomography. Treatment of KRG (p.o., 250 mg/kg of body weight/d) significantly preserved trabecular bone volume, trabecular number, structure model index, and bone mineral density of proximal tibia metaphysic, but did not alter the uterus weight of the mice. Serum ALP level was slightly reduced by KRG treatment. However, grip strength, mechanical property, and cortical bone architecture did not differ among the experimental groups. The results indicate that KRG can prevent radiation-induced bone loss in mice.

• The Journal of Advanced Prosthodontics
• /
• v.6 no.6
• /
• pp.512-520
• /
• 2014

PURPOSE. The purpose of this study was to assess the surface characteristics and the biocompatibility of zirconium (Zr) coating on Ti-6Al-4V alloy surface by radio frequency (RF) magnetron sputtering method. MATERIALS AND METHODS. The zirconium films were developed on Ti-6Al-4V discs using RF magnetron sputtering method. Surface profile, surface composition, surface roughness and surface energy were evaluated. Electrochemical test was performed to evaluate the corrosion behavior. Cell proliferation, alkaline phosphatase (ALP) activity and gene expression of mineralized matrix markers were measured. RESULTS. SEM and EDS analysis showed that zirconium deposition was performed successfully on Ti-6Al-4V alloy substrate. Ti-6Al-4V group and Zr-coating group showed no significant difference in surface roughness (P>.05). Surface energy was significantly higher in Zr-coating group than in Ti-6Al-4V group (P<.05). No difference in cell morphology was observed between Ti-6Al-4V group and Zr-coating group. Cell proliferation was higher in Zr-coating group than Ti-6Al-4V group at 1, 3 and 5 days (P<.05). Zr-coating group showed higher ALP activity level than Ti-6Al-4V group (P<.05). The mRNA expressions of bone sialoprotein (BSP) and osteocalcin (OCN) on Zr-coating group increased approximately 1.2-fold and 2.1-fold respectively, compared to that of Ti-6Al-4V group. CONCLUSION. These results suggest that zirconium coating on Ti-6Al-4V alloy could enhance the early osteoblast responses. This property could make non-toxic metal coatings on Ti-6Al-4V alloy suitable for orthopedic and dental implants.