• 한국미생물·생명공학회지
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• 제25권2호
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• pp.218-223
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• 1997

For the utilization of animal blood produced in slaughter for the cultivation of lactic acid bacteria, the nitrogen sources in a complex(MRS) medium were replaced by blood plasma proteins. Focusing the purpose on the industrial production of a probiotics, the hydrolytic activities of three industrially applicable proteases were compared for the effective digestion of the proteins, and Alcalase(the product of Novo Nordisk) was selected with comparatively high activity. The growth of Streptococcus thermophilus KCCM12020 was best among the four strains of lactic acid bacteria tested. With Alcalase-digested proteins in the medium, the growth rates and the final cell concentrations were higher than those with non-digested proteins. The cell mass produced in the medium containing blood proteins as nitrogen sources, $2.5{\times}10^9$ CFU/ml, was significantly high and about 70% of that in MRS medium, showing a great possibility for the utilization of animal blood proteins as economic nitrogen sources in the production of cell mass of lactic acid bacteria.

• The Korean Journal of Physiology and Pharmacology
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• 제28권5호
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• pp.435-447
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• 2024

Secretory proteins, including plasma membrane proteins, are generally known to be transported to the plasma membrane through the endoplasmic reticulum-to-Golgi pathway. However, recent studies have revealed that several plasma membrane proteins and cytosolic proteins lacking a signal peptide are released via an unconventional protein secretion (UcPS) route, bypassing the Golgi during their journey to the cell surface. For instance, transmembrane proteins such as the misfolded cystic fibrosis transmembrane conductance regulator (CFTR) protein and the Spike protein of coronaviruses have been observed to reach the cell surface through a UcPS pathway under cell stress conditions. Nevertheless, the precise mechanisms of the UcPS pathway, particularly the molecular machineries involving cytosolic motor proteins, remain largely unknown. In this study, we identified specific kinesins, namely KIF1A and KIF5A, along with cytoplasmic dynein, as critical players in the unconventional trafficking of CFTR and the SARS-CoV-2 Spike protein. Gene silencing results demonstrated that knockdown of KIF1A, KIF5A, and the KIF-associated adaptor protein SKIP, FYCO1 significantly reduced the UcPS of △F508-CFTR. Moreover, gene silencing of these motor proteins impeded the UcPS of the SARS-CoV-2 Spike protein. However, the same gene silencing did not affect the conventional Golgi-mediated cell surface trafficking of wild-type CFTR and Spike protein. These findings suggest that specific motor proteins, distinct from those involved in conventional trafficking, are implicated in the stress-induced UcPS of transmembrane proteins.

• Journal of Nutrition and Health
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• 제33권2호
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• pp.167-178
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• 2000

The effect of oral vitamin e (800IU/day) and C (500mg/day) supplementation for 10 days and/or smoking cessation for 5 days on oxidative damage to the red blood cells (RBC) of male smokers (22.2$\pm$0.2 years old) was studied. RBC were tested for their ability to protect against smoking-induced oxidative damage by measuring heme proteins (carboxyhemoglobin, hemoglobin, methemoglobin, oxyhemoglobin), hemolysis and thiobarbiturinc acid reactive substances (TBARS). Plasma levels of vitamin c, A, E, $\beta$-catotene, total cholesterol, glutamic pyruvic transaminase(GPT) and glutamic oxaloacetic transaminase(GOT) were also analyzed. In experiment one, a comparison was made of heme proteins and lipid damage to RBC, plasma antioxidant status (indexed by plasma levels of vitamin C, E, A and $\beta$-carotene) between smokers(n=56) and non-smokers (n=16). No differences were found in plasma antioxidant status, heme protein damage and TBARS concentration of RBC. In experiment two, 46 fasting male smokers from experiment one were divided into 4 groups. The groups were smoking with placebo group(SP, n=14), smoking cessation with vitamins supplementatin group (SV, n=13), smoking cessation with placebo group (NSP, n=9) and smoking cessation with vitamins supplementation group (NSV, n=10). After supplementing antioxidant vitamins, significant increases were seen in plasma vitamins supplementation group (NSV, n=10). After supplementing antioxidant vitamins, significant increases were seen plasma vitamin C (p<0.05) and vitamin E levels (p<0.05). The plasma vitamin E level was highest in the NSV group. Vitmain E and C supplementation provided some protection against heme proteins and lipid damage by lowering methemoglobin, hemolysis and TBARS concentration of RBC. Smoking cessation significantly decreased TBARS of RBC and plasma total cholesterol concentration. Supplementing vitamin E and C with smoking cessation considerably lowered plasma total cholesterol. These results point to a special association among smoking, oxidative damage and plasma antioxidant vitamin status. They indicate that increases in plasma antioxidant status can be detected after the supplementation of vitamin C and E and that smoking cessation had an additional effect on plasma vitamin E level. The present data suggest that improved antioxidant status induced by antioxidant supplementation or smoking cessation may help prevent oxidative damage in smokers.

• KSBB Journal
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• 제11권6호
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• pp.630-635
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• 1996

IgG와 알부민이 고스트 적혈구의 분산하에서 적 혈구의 세포막에 흡착되었다. 용액중 IgG의 농도가 0 O.1mg/mL일 때, 고스트 적혈구의 부피비가 5% 에 서 45%로 증가함에 따라 흡착으로 인한 용액중의 IgG의 농도감소는 14%에서 45%로 증가하였고, h hardened 고스트 척혈구 분산시가 고스트 적혈구 분산시보다 더 많은 단백질이 흡착되었다. 용액중 알부민의 농도가 0.075mg/mL일 때, 고스트 적헐구 의 부피가 5%에셔 70%로 증가함에 따라 용액중의 얄부민의 농도감소는 12% 에서 47%로 증가하였다. 흡착시간에 대한 IgG의 농도변화를 측정한 결과 단 백질의 흡착은 빠른 속도로 일어남을 알 수 있었고, 적혈구당 흡착된 단백질 분자수를 계산한 결과 용액 내 적혈구의 수가 증가할수록 적혈구당 흡착된 분자 수는 감소하였다.

• BMB Reports
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• 제31권4호
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• pp.364-369
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• 1998

Insect plasma protein is abundant in the hemolymph of holometabolous insect larvae and is used as a source of amino acids and energy for construction of adult structures during metamorphosis. In order to understand the mechanism of decomposition of larval plasma proteins by hemocyte protease, we tried to purify a cysteine protease from the hemocyte lysate by using Carbobenzoxy-L-Phenylalanyl-L-Arginine-4-Methyl-Coumaryl-7-Amide (Z-Phe-Arg-MCA) as substrate and to identify plasma proteins that are selectively susceptible to the purified protease. Here, we describe the purification and characterization of a cysteine protease that specifically hydrolyzes the plasma protein of the coleopteran insect, Tenebrio molitor, larvae. The molecular mass of this enzyme was 25 kDa, as determined by SDS-PAGE under reducing conditions. The amino acids sequence of its $NH_{2}-terminus$ was determined to be Leu-Pro-Gly-Gln-Ile-Asp-Trp-Arg-Asp-Lys-Gly. This sequence contained Pro, Asp, and Arg residues, conserved in many papain superfamily enzymes. The specific cysteine protease inhibitors, such as E-64 and leupetin, inhibited its hydrolytic activity. One plasma protein with a molecular mass of 48 kDa was selectively hydrolyzed within 3 h when the purified enzyme and plasma proteins were incubated in vitro. However, the 48 kDa protein was not hydrolyzed by the purified 25 kDa protease in the presence of E-64. Western blotting analysis at various developmental stages showed that the purified enzyme was detected at larvae, pupae, and adult stages, but not the embryo stage.

• Reproductive and Developmental Biology
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• 제34권3호
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• pp.235-240
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• 2010

This study was performed to comprehend the plasma proteins expressed specifically during early pregnancy in pregnant or non-pregnant Hanwoo using proteomic analysis technique. Plasma samples (0, 2, 3, 4, 7, and 11 weeks after AI) were obtained from pregnant (P, n=3) or non-pregnant (NP, n=4) Hanwoo, respectively. To evaluate proteins differentially expressed, 2-dimensional electrophoresis (2DE) was conducted. Normalized protein spots were selected for the significant expression variation deviated over two fold in its expression level between two groups; Molecular functions of the proteins were DNA binding, protein binding, hemoglobin binding, ferrochelatase and transporter activity and arylestera, respectively. According to western blotting, haptoglobin was specifically expressed only in NP group during early pregnancy; however, paraoxonase 1 was highly expressed in pregnant group. Based on these results, pregnancy was maintained successfully by the activation of specific plasma proteins associated with immune system and antioxidant regulation during early pregnancy in Hanwoo.

• 한국식품과학회지
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• 제30권4호
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• pp.988-991
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• 1998

소와 돼지 혈액중의 혈장 단백질을 분리하여, 성분 분석과 단백질 함량을 정량했으며 혈장단백질의 식품 기능성에 관하여서는 기존 유화제로서 활용되는 유청 단백질인 WPC와 비교하여 유화능력을 비교 검토하였다. FPLC 및 SDS-PAGE에 의한 성분분석 결과 혈장 알부민(SA)은 돈혈장단백질(PPP), 우혈장단백질(BPP), 유장단백질(WPC)의 순서로 많이 함유되었으며, 혈장 단백질에는 ${\beta}-globulin$분획(주로 transferrin)이 다량 포함되어 있고, 그외 미량성분(fibrinogen, immunoglobulin)들이 확인되었다. 단백질 함량은 BPP (85%) 및 PPP (82%) 모두 높은 함량을 보여 우수한 단백질원으로 이용 가능할 것으로 보였다. 유화력도 혈장단백질들이 단백질 농도 2% 이하에서는 WPC보다 높았으며, 4% 이상에서는 WPC보다는 약간 낮았다. 또한 염농도 및 pH 의존성에 관하여 유화력에 미치는 영향을 검토한 결과, PPP 및 BPP의 pH 영역에 대한 유화활성은 WPC와는 상이하게 산성쪽의 pH에서 염기성쪽보다 더 높은 활성을 보였으며, 염(NaCl)첨가로 인한 유화 활성의 영향은 WPC와 비교하여 pH 의존성이 상당히 높았으며, 특히 산성쪽에서 높은 활성을 나타냈다. 이상의 결과들을 살펴볼 때 소 및 돼지의 혈액에서 제조한 혈장단백질들은 우수한 유화특성을 갖는 식품소재로 확인되었다.

• Journal of Animal Science and Technology
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• 제58권8호
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• pp.31.1-31.8
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• 2016

Background: Plasma protein hydrolysates have been shown to possess antioxidant activity. However, no report has yet to examine the antioxidant effects of injection of plasma protein hydrolysates on meat quality. Therefore, in this study, the effects of injection of hydrolysis plasma protein solution on meat quality and storability were investigated in porcine M. longissimus lumborum. Methods: Twelve pigs were randomly selected at a commercial slaughter plant and harvested. Dissected loins were injected with one of five solutions: C- control (untreated), T1- 10 mM phosphate buffer solution (PBS), T2- 10 mM PBS with 0.01 % butylated hydroxytoluene, T3- 10 mM PBS with 5 % plasma proteins, and T4- 10 mM PBS with 5 % hydrolysis plasma proteins. Results: T3 and T4 induced greater reduction in protein content of the loin muscle than other treatments. T2 resulted in the lowest pH as well as highest cooking loss. After a storage period of 3-7 days, both lightness and redness of meat were unaffected by all injection treatments. However, yellowness was significantly elevated by treatment with T4 relative to the control. T4 also resulted in the lowest shear force (a measure of meat toughness), suggesting improvement of texture or tenderness. Further, T4 resulted in the most stable TBARS values during storage, indicating that this treatment might retard rancidity in meat. Conclusion: Injection of porcine M. longissimus lumborum with hydrolysis plasma protein solution could improve overall pork quality, including tenderness and storability.

• BMB Reports
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• 제37권3호
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• pp.362-369
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• 2004

The human folate receptor (hFR) is a glycosylphosphatidylinositol (GPI) linked plasma membrane protein that mediates delivery of folates into cells. We studied the sorting of the hFR using transfection of the hFR cDNA into MDCK cells. MDCK cells are polarized epithelial cells that preferentially sort GPI-linked proteins to their apical membrane. Unlike other GPI-tailed proteins, we found that in MDCK cells, hFR is functional on both the apical and basolateral surfaces. We verified that the same hFR cDNA that transfected into CHO cells produces the hFR protein that is GPI-linked. We also measured the hFR expression on the plasma membrane of type III paroxysmal nocturnal hemoglobinuria (PNH) human erythrocytes. PNH is a disease that is characterized by the inability of cells to express membrane proteins requiring a GPI anchor. Despite this defect, and different from other GPI-tailed proteins, we found similar levels of hFR in normal and type III PNH human erythrocytes. The results suggest the hypothesis that there may be multiple mechanisms for targeting hFR to the plasma membrane.

• Parasites, Hosts and Diseases
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• 제48권3호
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• pp.203-211
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• 2010

Advancements in the field of proteomics have provided great opportunities for the development of diagnostic and therapeutic tools against human diseases. In this study, we analyzed haptoglobin and amyloid A protein levels of vivax malaria patients with combinations of depletion of the abundant plasma proteins, 2-dimensional gel electrophoresis (2-DE), image analysis, and mass spectrometry in the plasma between normal healthy donors and vivax malaria patients. The results showed that the expression level of haptoglobin had become significantly lower or undetectable in the plasma of vivax malaria patients due to proteolytic cleavage when compared to healthy donors on 2-DE gels. Meanwhile, serum amyloid A protein was significantly increased in vivax malaria patient's plasma with high statistical values. These 2 proteins are common acute phase reactants and further large scale evaluation with a larger number of patient's will be necessary to establish the possible clinical meaning of the existential changes of these proteins in vivax malaria patients. However, our proteomic analysis suggests the feasible values of some plasma proteins, such as haptoglobin and serum amyloid A, as associating factor candidates for vivax malaria.