• Title/Summary/Keyword: C-반응성 단백질

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Protein kinase C와 이와 관련된 단백질 연구

  • 이재란;김진한;최명언
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.135-135
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    • 1993
  • 신호전달과정에 중요한 역할을 하고 있는 다기능 serinei/threonine 단백질인산화효소인 protein kinase C(PKC)의 연구를 위해 이 효소의 정제를 뇌에서 착수하였다 PKC의 활성측정을 myelin basic protein을 기질로 하여 20 mM Tris 완충용액 PH 7.5, 0.15 mM [${\gamma}$-$^{32}$P]ATP(3 $\times$ $10^{5}$ cpm), 0.1 mM $Ca^{2+}$, 10$\mu\textrm{g}$ phosphatidylserine과 2$\mu\textrm{g}$ diolein을 넣어 반응시켰다. 반응은 TCA로 정지시킨 후 방사성 단백질을 Millipore filter paper로 걸러 섬광 계수기로 읽었다. Cytosol PKC의 정제과정은 첫 단계에서 DEAE-cellulose를 사용하였으며, phenyl sepharose CL-4B와 protamine agarose를 연속적으로 이용하여 800배의 정제에 성공했다. SDS-PACE 상에서 80 kD로 나타났으며 순도는 95 % 이상이였다. 이를 이용 PKC의 각종 기질 연구에 착수하기 시작했으며, 이중 MBP의 인산화연구를 통한 myelin의 안정성과 MBP와의 구조 관계가 일부 수행되고 있다 연차적으로 PKC와 이와 관련된 단백질의 특성을 살피기 위해 뇌의 PKC 기질 중 cold stress를 통해 환경에 민감한 것을 찾고 있으며, 현재 autoradiography를 이용해 80 kD, 54 kD, 49 kD와 35 kD의 단백질이 연구대상이 되고있다. 그 중 49 kD는 B-50(또는 GAP43, neuromodulin이라고도 함)일 가능성이 높아 이 단백질 조절과 PKC 활성화 사이의 관계 정립이 흥미로운 과제로 대두되고 있다.다.

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Immunological Analysis of Endotoxin Proteins Produced by Bacillus thuringiensis serovar. kurstaki HD1 and HA73 (Bacillus thuringiensis serovar. kurstaki HD1과 HD 73이 생산하는 내독소 단백질의 면역학적 분석)

  • 오상수;이영종;김창규;구본성;김종배;이형환
    • Microbiology and Biotechnology Letters
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    • v.16 no.2
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    • pp.168-173
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    • 1988
  • Immunological analysis between endotoxin proteins produced by Bacillus thuringiensis serovar. kurstaki HD1 and HD73 have been investigated by using polyclonal antibodies. The antisera against the endotoxin proteins were prepared from rabbits injected with the endotoxin protein antigens. When about 2mg/$m\ell$ of the antigens were injected for 7 times, the titers were highest. The stability of the antigens was reduced to about 50% after 9 days incubation at 4$^{\circ}C$. The sensitibity of endotoxin protein from B. thuringiensis HD1 and HD73 by indirect ELISA was 50ng/$m\ell$ and 400ng/$m\ell$, respectively. The cross reaction of antiserum appeared that anti-HD1 partialy reacted with crystal protein but anti-HD73 reacted with HD1 endotoxin about 100%.

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The Clinical Value of Intima-media Thickness in Patients with Stable Coronary Artery Disease and C-reactive Protein within Normal Limits (경동맥 내중막 두께가 정상 C-반응성 단백질 농도를 가진 안정형 협심증 환자에서 가지는 임상적 의의)

  • Kwon, Jong-Bum;Park, Kuhn;Choi, Si-Young;Park, Chan-Beom;Kim, Yong-Hwan;Her, Sung-Ho;Park, Mahn-Won;Lee, Jong-Ho
    • Journal of Chest Surgery
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    • v.43 no.5
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    • pp.482-489
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    • 2010
  • Background: The purpose of this study was to determine whether hand-measured carotid intima-media thickness (IMT) was a reliable and simple tool that could be used to evaluate the severity of stable coronary artery disease (CAD) with normal high-sensitivity C-reactive protein (CRP). Material and Method: We examined left carotid IMTs in 688 patients with stable CAD and excluded 121 patients with abnormally elevated hs-CRP ($\geq$0.5 mg/dL). We sought to determine the association between risk factors and the severity of stable CAD and carotid IMT in 567 patients who were divided into lower and higher groups based on the median value. Patients underwent a coronary artery angiogram and had at least significant stenosis (>50% of the original luminal diameter involving one or more major coronary artery). Result: Of the 567 patients, 300 (52.9%) were in the lower IMT group and 267 (47.1%) were in the higher IMT group; the median value was 0.76 mm. By multivariable logistic regression analysis, the following groups were different: older age (~49 vs. 70~ years, respectively; OR=6.552), high FBS (~99 vs. 120~ mg/dL; OR=1.713) and severity of CAD (1 vessel vs. 2 vessel disease; OR=1.711, 1 vessel vs. 3 vessel disease; OR=1.714). Conclusion: We conclude that there are correlations between increased carotid IMT and severity of CAD in stable angina patients with normal CRP levels.

The Effect of Sugar Addition and Heat Treatment on the Myofibrillar Protein Extractability (당첨가 및 가열처리가 근원섬유단백질의 추출성에 미치는 영향)

  • Yang, Jong-Beom;Kim, Chang-Han
    • Korean Journal of Food Science and Technology
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    • v.22 no.4
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    • pp.466-472
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    • 1990
  • The effect of sugar addition and heat treatment on the myofibrillar protein extractability was studied. Maillard reaction was dependent on heating time significantly and glucose revealed the highest reactivity for Maillard reaction. The extractability of myofibrillar proteins was lowest in case of glucose addition and decreased according to increasing of heating time, Higher extractability was resulted in by digestion of myofibrillar proteins with enzymes after sugar addition and heat treatment than the undigested samples, as the sample was digested with trypsin that is the highest. And by digestion with trypsin, chymotrypsin and peptidase at a time the extractability of meat proteins increased remarkably.

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Immune Response of Bacterial Proteins of Staphylococcus intermedius from Canine Atopic Dermatitis (개의 아토피성 피부염에서 분리한 Staphylococcus intermedius 균의 세균단백질의 면역반응)

  • Park, Hee-myung
    • Journal of Veterinary Clinics
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    • v.21 no.1
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    • pp.20-22
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    • 2004
  • Bacterial infection of canine atopic dermatitis is largely caused by Staphylococcus intermedius and may be a superficial or deep pyoderma. The Purpose of this study was to identify the major proteins of S. intermedius cell surface components in humoral immune response of atopic dermatitis dog. Sera samples were obtained from dogs with atopic dermatitis and superficial pyoderma referred to the Veterinary Medical Teaching Hospital, Konkuk University. An isolate of S. intermedius from a clinical case of canine atopic dermatitis was cultured in brain heart infusion broth overnight at $37^{\circ}C$ in aerobic conditions on an orbital shaker. Following culture, Staphylococci were harvested by centrifugation, washed in PBS, and resuspended in PBS containing lysostaphin. The soluble components were separated by centrifugation and were collected. The soluble extract of S. intermedius was separated by sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE). The proteins were electrophoretically transferred onto nitrocellulose membrane. Western blotting for the specificity of serum IgG antistaphylococcal antibody was performed with anti-dog-IgG and sera obtained from an atopic dermatitis case and a normal dog. The molecular masses of four major proteins of S. intermedius recognized by serum obtained from an atopic dermatitis case were 18, 31, 75, and 110 kDa as determined by Western blot analysis. The present study indicates that most dogs of S. intermedius infection with atopic dermatitis could have a significant humoral immune response to bacterial proteins of the causative organism.

Process Optimization of Peptides Production from Protein of Crab (Ovalipes punctatus) and Its Antioxidant Capacity Analysis (꽃게(Ovalipes punctatus) 단백질 유래 항산화 기능성 펩타이드 제조 최적공정 확립 및 이화학적 특성)

  • Ha, Yoo Jin;Kim, Do Hyun;Lee, Byung Hee;Yoo, Sun Kyun
    • Journal of the Korean Applied Science and Technology
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    • v.35 no.2
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    • pp.367-377
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    • 2018
  • Swimming crab(Ovalipes punctatus) is produced in Korea and utilized as semi-processed food at streamed cooked state. Recently, protein hydrolysates have been known as having function such as antioxidant, suppression of hypertension, immunodulatory, alleviation of pain, and antimicrobial activity. This research was investigated to find the functional antioxidant from crab hydrolysates. To fine optimal protease enzyme, alcalase, bromelain, flavourzyme, neutrase, papain, and protamex were selected to evaluate the DPPH radical scavenging activity and finally bromelain to show the best activity was selected. The molecular weight of bromelain hydrolysates were distributed with range from 500 to 3,200 Da and 7 different molecules or more. The amino acids related to antioxidant capacity was about 42.54%. The processes optimization study used was the response surface methodology. The ranges of processes were the reaction temperature of 40 to $60^{\circ}C$, pH 6 to 8, and enzyme concentration 1 to 3%(w/v). As a result, the optimization of process was determined at temperature of $55^{\circ}C$, pH of 6.5, and enzyme concentration of 3%(w/v). In these conditions, degree of hydrolysates were maximum 71.60%. Therefore, we expect that those products are useful as functional food ingredients.

Involvement of Oxidative Stress in Formaldehyde-induced Apoptosis in Cultured Lung Macrophage Cells (폐 대식세포주에서 포름알데히드에 의한 세포 사멸 효과에 대한 산화성 스트레스 관련성)

  • Park, Soo-Hyun
    • Korean Journal of Environmental Agriculture
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    • v.28 no.3
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    • pp.295-300
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    • 2009
  • Formaldehyde (FA) is an important irritant compound in pesticide to induce asthma and allergy in respiratory system. Alveolar macrophage is also an pivotal cell in the immune response of respiratory system. However, the effect of FA in macrophage cell viability has not been elucidated. Thus, this study was conducted to investigate the effect of FA on apoptosis in Raw 264.7 cells, alveolar macrophage cell line. In this study, FA decreased cell viability of lung alveolar macrophage cells in a dose-dependent manner (>$100{\mu}M$). FA-induced decrease of cell viability was blocked by the treatment of antioxidants (vitamin C, NAC, and catalase). Indeed, FA induced lipid peroxide formation in Raw 264.7 cells. FA decreased Bcl-2 expression but increased Bax expression in lung alveloar macrophage cells. In addition, FA also increased the cleaved form of caspase-3. In conclusion, FA induced apoptosis via oxidative stress in cultured Raw 264.7 cells.

Dephosphorylation Study of Phosphorylated Myelin Basic Protein: A Model Substrate for Protein Phosphatase (인산화된 신경수초 염기성 단백질의 탈인산화 연구: 단백질 탈인산화 효소의 기질 모델)

  • Kim, Jin Hahn;Choi, Myung Un
    • Journal of the Korean Chemical Society
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    • v.41 no.4
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    • pp.205-209
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    • 1997
  • The site specificity of dephosphorylation of myelin basic protein(MBP) was studied in vitro. To assign amino acid site of dephosphorylation, MBP was phosphorylated by protein kinase C(PKC) and dephosphorylated by protein phosphatase PP2A. The phosphorylated MBP was digested by trypsine and the digested peptides were separated by a reverse phase HPLC chromatography. The radioactivity of each fraction was counted and partially sequenced. Seven radioactive peptides were observed and $Ser^{55}$ in the second peak($P_2$) shows the best susceptibility for the phosphorylation. However in the dephosphorylation, the fifth peak($P_5$) appeared to release it's phosphate group most rapidly. This result demonstrates that MBP is a suitable substrate for protein phosphatase.

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Dose-dependent Effects of Dietary Folate on Aortic Relaxation and Hepatic C-reactive protein Levels in C57BL/6 Mice (C57BL/6 쥐의 대동맥 이완과 간 C반응단백질 수준에 미치는 섭취엽산의 용량의존 효과)

  • Kong, Eunhee;Hasan, Syeda T.;Jang, Hyeran;Zimmerly, Ella M.;Choi, Sang-Woon;Meydani, Mohsen
    • Journal of Life Science
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    • v.25 no.8
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    • pp.889-895
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    • 2015
  • Endothelial dysfunction is an initial step in atherosclerosis. B vitamins (B6, B12, and folate) are important contributing factors to vascular homeostasis. Deficiencies in these B vitamins induce cardiovascular diseases by altering vascular homeostasis. Folate plays important roles in nitric oxide homeostasis in the endothelium. To determine the dose-dependent effect of dietary folate on atherosclerosis, we studied aortic relaxation and hepatic C-reactive protein (CRP) levels in C57BL/6 mice. In this study, a total of 54 male C57BL/6, 8-wk old mice were split into 2 dietary groups (control and Western style diet). Each diet group was divided into 3 subgroups according to dietary folate dosage (0.2, 2, and 8 mg/kg). After 18 months, the relaxation response seen in aortic rings from mice fed 0.2 or 2 mg folate/kg in both diet groups. However, the aortic relaxation response was not seen and no differences were observed in mice fed 8mg folate/kg in either diet group (p<0.05). Hepatic CRP levels at all folate dosages (0.2, 2, 8 mg folate/kg) were higher in the groups fed a Western style diet than in mice fed a control diet (p=0.035). CRP levels were lower in mice fed 0.2 mg folate/kg than in mice fed 2 or 8 mg folate/kg in both diet groups (p<0.05). These results indicate that in C57BL/6 mice 0.2 mg folate/kg may be enough to prevent atherosclerosis by inducing the relaxation responses of the aorta and by reducing levels of hepatic CRP, regardless of dietary style.

Campylobacter jejuni 의 열충격 반응과 그유전자에 관한 연구

  • 김치경;임채일;이길재
    • Korean Journal of Microbiology
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    • v.30 no.3
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    • pp.232-238
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    • 1992
  • Canz~~j~lohuc;tc.~jurn i werc studied for their heat shock responses at several elevated temperatures and their heat shock genes were detected by the technique of Southern hybridization. (.. ,jc\ulcorneruni sy~>thesized the major heat shock proteins of hsp90. hsphh. and hsphO at 48$^{\circ}$C . ant1 their w~u.ival rates were maintained as the same level at optimal temperature. '1-hc heat shock genes in chromosome of C ,jc:jutii werc determined to be homologous to the heat shock genes or E. t,oli. by showing strong signals in Southern hybridization analysis using clnaK and groESL- as DNA probe But the restriction sites for thc fragmcnts including heat shock genes were different betueen E. c,oli and C ,jtjuni.

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