• 제목/요약/키워드: modified protein

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Pepsin으로 수식된 정어리 myofibrillar protein의 특성 (Physicochemical and Functional Properties of Pepsin-modified Myofibrillar protein from Sardine, Sardinops melanostica)

  • 김병묵;김병열
    • 한국식품과학회지
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    • 제26권2호
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    • pp.110-116
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    • 1994
  • 효소적수식에 따른 정어리 myofibrillar protein의 특성변화를 연구하기 위하여 1/100량의 pepsin으로 $37^{\circ}C$. pH 1.65에서 각각 1, 4, 8, 12, 24시간 부분가수분해시킨 후 단백질의 물리화학적 및 기능적 특성을 검토하였다. 정어리 myofibrillar protein은 pepsin에 의하여 가수분해 1시간까지는 직선적으로 가수분해율이 증가되어 약 30%의 가수분해율을 나타내었고 가수분해 4시간에는 약 40%의 가수분해율을 나타내었으나, 그 이후에는 더 이상 가수분해되지 않아 가수분해 24시간에도 가수분해율은 크게 증가되지 않았다. 전기영동상은 분자량 40,000정도의 굵은 band는 큰 변화를 나타내지 않고 비교적 분자량이 큰 것으로 보이는 상층부의 몇몇 엷은 band와 분자량이 작은 하층부의 band가 가수분해가 진행되면서 소실되었다. 중간부위에 위치하는 몇몇 엷은 band는 가수분해시간이 지나면서 하나로 뭉쳐지는 듯한 경향을 나타내었다. Gel여과 pattern은 두개의 큰 peak와 세개의 작은 peak를 나타내 주고 있는데 가수분해 1시간에 작은 peak는 모두 사라지고 앞쪽의 큰 peak는 가수분해시간이 지남에 따라 점차 작아지고 뒷쪽의 큰 peak는 가수분해시간이 지남에 따라 점차 약간씩 뒤로 밀려가는 경항을 나타내었다. 유화활성과 유화용량은 pepsin 가수분해에 의하여 모두 감소되었으며 특히 유화용량은 가수분해 시간이 경과되면서 현저히 감소되어 가수분해 24시간에는 약 59%로 저하되었다. 유화활성은 가수분해기간중 큰 변화를 나타내지 않아 가수분해 24시간에도 약 94%의 유화활성이 유지되었다. 기포력은 크게 증가되어 가수분해 24시간에는 대조구의 1.9배나 증가되었다. 거품안 정성은 반대로 감소되어 가수분해 8시간에는 약 0.6배로 감소되었으나 그 이후에는 다시 증가되어 가수분해 24시간에는 약 0.8배로 감소율이 둔화되었다. 열응고성은 pepsin수식에 의하여 0.55 내지 0.69배로 감소되었으며 점도는 가수분해시간이 길어지면서 점차 증대되어 가수분해 24시간에는 약 1.36배 증대되었다. 흡수율은 현저히 떨어져 가수분해 1시간에 약 0.32배로 감소되었고 용해도는 현저히 커져 가수분해 1시간에 약 1.46배로 증대되었다. 그러나 그 이후에는 흡수율이나 용해도 모두 더 이상 큰 변화를 나타내지 않았다. 용해도의 pH의존성은, pH 5와 9부근에서 다소 낮은 용해도를 나타내었으며 pepsin수식에 의하여 이러한 경향은 가수분해 초기인 1시간에는 더욱 뚜렸이 나타났다가 가수분해 후기인 24시간에는 크게 둔화되는 양상을 나타내었다.

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가소제 및 가교제에 의해 개질된 대두단백질의 특성 (Characteristics of Soybean Protein Resin Modified by Plasticizers and Cross-Linking Agents)

  • 최한나;이태상;양지우;이승구
    • 접착 및 계면
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    • 제12권2호
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    • pp.73-80
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    • 2011
  • 식물성 고분자인 대두단백질을 기반으로 하는 환경친화성 고분자 신소재에 관한 연구를 위해 가소제(1,3-propandiol, glycerol) 및 가교제(glutaraldehyde, epichlorohydrin, glyoxal, urea)에 의한 대두단백질 수지의 열적 특성을 TGA를 이용하여 분석하였고, 기계적 특성 분석과 SEM을 통하여 파단면을 관찰하였다. 그 결과, 가소제인 1,3-propandiol과 glycerol을 SPI (대두단백질)에 첨가함으로써 수지의 유연성이 증가하였고, 1,3-propandiol에 비하여 glycerol의 가소화 효과가 상대적으로 크게 나타났으며, 가교제인 glycerol, epichlorohydrin, glyoxal의 적용으로 첨가량이 증가할수록 대두단백질 수지의 강도와 열안정성이 증가하는 반면, urea의 경우, 대두단백과의 가교가 용이하지 않아 열안정성이 오히려 낮아지고, 강도가 감소함을 알 수 있었다.

Single-Protein Molecular Interactions on Polymer-Modified Glass Substrates for Nanoarray Chip Application Using Dual-Color TIRFM

  • Kim, Dae-Kwang;Lee, Hee-Gu;Jung, Hyung-Il;Kang, Seong-Ho
    • Bulletin of the Korean Chemical Society
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    • 제28권5호
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    • pp.783-790
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    • 2007
  • The immobilization of proteins and their molecular interactions on various polymer-modified glass substrates [i.e. 3-aminopropyltriethoxysilane (APTS), 3-glycidoxypropyltrimethoxysilane (GPTS), poly (ethylene glycol) diacrylate (PEG-DA), chitosan (CHI), glutaraldehyde (GA), 3-(trichlorosilyl)propyl methacrylate (TPM), 3'-mercaptopropyltrimethoxysilane (MPTMS), glycidyl methacrylate (GMA) and poly-l-lysine (PL).] for potential applications in a nanoarray protein chip at the single-molecule level was evaluated using prismtype dual-color total internal reflection fluorescence microscopy (dual-color TIRFM). A dual-color TIRF microscope, which contained two individual laser beams and a single high-sensitivity camera, was used for the rapid and simultaneous dual-color detection of the interactions and colocalization of different proteins labeled with different fluorescent dyes such as Alexa Fluor® 488, Qdot® 525 and Alexa Fluor® 633. Most of the polymer-modified glass substrates showed good stability and a relative high signal-to-noise (S/N) ratio over a 40-day period after making the substrates. The GPTS/CHI/GA-modified glass substrate showed a 13.5-56.3% higher relative S/N ratio than the other substrates. 1% Top-Block in 10 mM phosphate buffered saline (pH 7.4) showed a 99.2% increase in the blocking effect of non-specific adsorption. These results show that dual-color TIRFM is a powerful methodology for detecting proteins at the single-molecule level with potential applications in nanoarray chips or nano-biosensors.

골감소증을 동반한 지속성 복막투석환자의 다빈도섭취 음식조사 및 골밀도 개선을 위한 레시피 개발 (Frequently Consumed Dishes and Development of Recipes to Improve Bone Mineral densities in Continuous Ambulatory Peritoneal Dialysis Patients with Osteopenia)

  • 박진경;손숙미
    • 대한영양사협회학술지
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    • 제12권4호
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    • pp.411-431
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    • 2006
  • The purpose of this study was to assess the frequently consumed dish consumption frequencies of continuous ambulatory peritoneal dialysis(CAPD) patients with osteopenia and develop recipes to improve bone mineral density of CAPD. The subjects were 96 CAPD patients with osteopenia(male 39, female 57) (osteopenia group) and 45 CAPD patients with normal BMD(male 24, female 21), matched with key variables(normal group). Fifty dishes(foods) that most frequently consumed were determined and food consumption frequency for each dish(food) for two groups were compared. Osteopenia group showed lower consumption frequency for ice-cream but higher frequency in apple. Of the 50 most frequently consumed dishes(foods), 20 dishes assessed as safe and recommendable for CAPD patients with osteopenia based on the contents of protein and mineral were selected : white boiled rice, white gruel, beef soup, steamed cabbage, roasted dried laver, fried egg, roasted bean-curd, cooked and seasoned bean sprouts, corn-starch jelly, cheese, ice-cream, orange juice, apple, grape, peach, peanut, raw lettuce, raw cucumber, and injulmi rice cake. wenty eight new dishes with modified recipes were developed for CAPD patients. Protein and mineral contents were analyzed for frequently consumed 17 dishes, assessed as modification of recipes are needed. The recipes were modified to decrease P, Na and K contents and to increase protein and Ca contents. Twenty dishes(foods) selected as having reasonable protein and mineral contents ratio or 28 newly developed dishes modified with protein and mineral contents or ratio would be helpful for nutrition education or counseling for CAPD patients with osteopenia. Dishes(foods) suggested in this study would also be useful for all CAPD patients for preventing osteoporosis.

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MUSCLE PROTEIN SYNTHESIS IN VITRO IN CHICKS FED A LOW-PROTEIN DIET

  • Kita, K.;Kuzuya, Y.;Matsunami, S.;Okumura, J.
    • Asian-Australasian Journal of Animal Sciences
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    • 제9권2호
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    • pp.171-174
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    • 1996
  • Muscle protein synthesis in vitro was measured in chicks fed low-protein(10% CP) and control(20% CP) diets. Right leg muscles (M. gastrocnemius) were mounted on a support made of stainless steel to stretch in constant tension, whereas left leg muscles were unmounted. Both leg muscles were incubated in Dulbecco's modified Eagle's medium including L-[$4-^3H$] phenylalanine for 60 min to measure in vitro protein synthesis. There was no significant difference in fractional synthesis rate(FSR) of muscle protein between both dietary protein levels, whereas FSR with stretch in constant tension was significantly higher than that without constant tension due to an increase in the absolute synthesis rate(ASR) per unit RNA(the efficiency of RNA to synthesize protein). The ASR of muscle protein in chicks fed the control diet was significantly higher than that in the low-protein diet group.

Salt-Induced Protein Precipitation in Aqueous Solution: Single and Binary Protein Systems

  • Kim, Sang-Gon;Bae, Young-Chan
    • Macromolecular Research
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    • 제11권1호
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    • pp.53-61
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    • 2003
  • A molecular-thermodynamic model is developed for the salt-induced protein precipitation. The protein molecules interact through four intermolecular potentials. An equation of state is derived based on the statistical mechanical perturbation theory with the modified Chiew's equation for the fluid phase, Young's equation for the solid phase as the reference system and a perturbation based on the protein-protein effective two body potential. The equation of state provides an expression for the chemical potential of the protein. In a single protein system, the phase separation is represented by fluid-fluid equilibria. The precipitation behaviors are simulated with the partition coefficient at various salt concentrations and degree of pre-aggregation effect for the protein particles. In a binary protein system, we regard the system as a fluid-solid phase equilibrium. At equilibrium, we compute the reduced osmotic pressure-composition diagram in the diverse protein size difference and salt concentrations.

Regulation of Protein Degradation by Proteasomes in Cancer

  • Jang, Ho Hee
    • Journal of Cancer Prevention
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    • 제23권4호
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    • pp.153-161
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    • 2018
  • Imbalance of protein homeostasis (proteostasis) is known to cause cellular malfunction, cell death, and diseases. Elaborate regulation of protein synthesis and degradation is one of the important processes in maintaining normal cellular functions. Protein degradation pathways in eukaryotes are largely divided into proteasome-mediated degradation and lysosome-mediated degradation. Proteasome is a multisubunit complex that selectively degrades 80% to 90% of cellular proteins. Proteasome-mediated degradation can be divided into 26S proteasome (20S proteasome + 19S regulatory particle) and free 20S proteasome degradation. In 1980, it was discovered that during ubiquitination process, wherein ubiquitin binds to a substrate protein in an ATP-dependent manner, ubiquitin acts as a degrading signal to degrade the substrate protein via proteasome. Conversely, 20S proteasome degrades the substrate protein without using ATP or ubiquitin because it recognizes the oxidized and structurally modified hydrophobic patch of the substrate protein. To date, most studies have focused on protein degradation via 26S proteasome. This review describes the 26S/20S proteasomal pathway of protein degradation and discusses the potential of proteasome as therapeutic targets for cancer treatment as well as against diseases caused by abnormalities in the proteolytic system.

Comparative Studies of Protein Modification Mediated by Fenton-like Reactions of Iron, Hematin, and Hemoglobin: Generation of Different Reactive Oxidizing Species

  • Kim, Young-Myeong;Kim, Sung-Soo;Kang, Gu;Yoo, Yeong-Min;Kim, Ki-Mo;Lee, Mi-Eun;Han, Jeong-A;Hong, Sun-Joo
    • BMB Reports
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    • 제31권2호
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    • pp.161-169
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    • 1998
  • TThe reactive oxygen species oxidatively modify the biological macromolecules, including proteins, lipids, and nucleic acids. Iron- and heme-mediated Fenton-like reactions produce different pro-oxidants. However, these reactive products have not been clearly characterized. We examined the nature of the oxidizing species from the different iron sources by measuring oxidative protein modification and spectroscopic study. Hemoglobin (Hb) and methemoglobin (metHb) were oxidatively modified in $O{\array-\\\dot{2}}$ and $H_{2}O_{2}$ generating systems. Globin and bovine serum albumin (BSA) were also modified by iron, iron-EDTA, hematin, and Hb in an $O{\array-\\\dot{2}}$ generating system. In a $H_{2}O_{2}$ generating system, the iron- and iron-EDTA-mediated protein modifications were markedly reduced while the Hb-and hematin-mediated modifications were slightly increased. In the $O{\array-\\\dot{2}}$ generating system, the iron- and iron-EDTA-mediated protein modifications were strongly inhibited by superoxide dismutase (SOD) or catalase, but heme- and Hb-mediated protein modifications were inhibited only by catalase and slightly increased by SOD. Mannitol, 5,5-dimethyl-l-pyrroline-N-oxide (DMPO), deoxyribose, and thiourea inhibited the iron-EDTA-mediated protein modification. Mannitol and DMPO, however, did not exhibit significant inhibition in the hematin-mediated modification. Desferrioxamine (DFO) inhibited protein modification mediated by iron, but cyanide and azide did not, while the hematin-mediated protein modification was inhibited by cyanide and azide, but not significantly by DFO. The protein-modified products by iron and heme were different. ESR and UV-visible spectroscopy detected the DMPO spin adduct of the hydroxyl radical and ferryl ion generated from iron-EDTA and metHb, respectively. These results led us to conclude that the main oxidizing species are hydroxyl radical in the iron-EDTA type and the ferry I ion in the hematin type, the latter being more effective for protein modification.

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참돔(Pagrus major)의 혈장 난황단백전구체에 대한 효소면역측정법과 난소성숙의 조기판정 (Enzymeimmunoassay for the Plasma Vitellogenin and Early Determination of Ovarian Maturation in Red Seabream, Pagrus major)

  • 한창희;양문호;백재민;임상구;김광현
    • 한국양식학회지
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    • 제8권1호
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    • pp.1-19
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    • 1995
  • 난황형성 중인 암 참돔의 혈청 내에는 암 특이혈청단백질이 존재하고 있음이 Ouchterlony의 면역확산 검정과 면역 전기영동에 의하여 밝혀졌으며, 이러한 암 특이혈청단백질은 암 $\cdot$수 혈청과 난추출액의 면역학적인 조사에 의해 난황단백 전구체임이 밝혀졌다. 또한 난추출액으로부터 정제된 난황단백질은 난황전구체와 공통의 항원성이 있음도 면역학적인 조사에 의하여 알 수 있었다. 변형한 효소면역측정법의 타당성을 조사하였다. 일정 비율로 희석한 성숙중인 암 혈청의 흡광도 곡선은 난황단백질의 표준곡선과 평행하였다 본 효소면역측정법의 평균 회수율은 $109\pm5.6\%$이었으며, $10\%$ 이내의 assay내 변동계수 범위는 $31\~1,000ng/ml$이었다. 생식소의 성숙 단계에 대한 혈장내의 난황단백전구체의 양과의 관계를 조사하기 위하여 휴지기후기(1월)부터 난황형성기(4월)까지 난황단백질에 대한 항체를 이용한 효소면역측정법에 의하여 혈장 난황단백전구체의 양을 측정하였다. 난황단백전구체의 양은 난황형성 전기인 2월부터 증가하기 시작하여, 난황형성기(3월부터 4월) 동안에는 난이 성장하면서 계속 증가하였다 2월부터 혈장내의 난황단백전구체의 양이 암$\cdot$수에 따라 현저하게 차이가 났다. 따라서 참돔 친어의 성 구분은 효소면역 측정법에 의하여 2월부터는 명확히 할 수 있다

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