• Title/Summary/Keyword: Leu-7

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Characterization of an Iron- and Manganese-containing Superoxide Dismutase from Methylobacillus Sp. Strain SK1 DSM 8269

  • Seo, Sung Nam;Lee, Jae Ho;Kim, Young Min
    • Molecules and Cells
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    • v.23 no.3
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    • pp.370-378
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    • 2007
  • A superoxide dismutase was purified 62-fold in seven steps to homogeneity from Methylobacillus sp. strain SK1, an obligate methanol-oxidizing bacterium, with a yield of 9.6%. The final specific activity was 4,831 units per milligram protein as determined by an assay based on a 50% decrease in the rate of cytochrome c reduction. The molecular weight of the native enzyme was estimated to be 44,000. Sodium dodecyl sulfate gel electrophoresis revealed two identical subunits of molecular weight 23,100. The isoelectric point of the purified enzyme was found to be 4.4. Maximum activity of the enzyme was measured at pH 8. The enzyme was stable at pH range from 6 to 8 and at high temperature. The enzyme showed an absorption peak at 280 nm with a shoulder at 292 nm. Hydrogen peroxide and sodium azide, but not sodium cyanide, was found to inhibit the purified enzyme. The enzyme activity in cell-free extracts prepared from cells grown in manganese-rich medium, however, was not inhibited by hydrogen peroxide but inhibited by sodium azide. The activity in cell extracts from cells grown in iron-rich medium was found to be highly sensitive to hydrogen peroxide and sodium azide. One mol of native enzyme was found to contain 1.1 g-atom of iron and 0.7 g-atom of manganese. The N-terminal amino acid sequence of the purified enzyme was Ala-Tyr-Thr-Leu-Pro-Pro-Leu-Asn-Tyr-Ala-Tyr. The superoxide dismutase of Methylobacillus sp. strain SK1 was found to have antigenic sites identical to those of Methylobacillus glycogenes enzyme. The enzyme, however, shared no antigenic sites with Mycobacterium sp. strain JC1, Methylovorus sp. strain SS1, Methylobacterium sp. strain SY1, and Methylosinus trichosproium enzymes.

Synthesis, Radiolabeling and Gastrin Releasing Peptide Receptor Binding Affinity of a Novel Bombesin Antagonist-Based Peptide, DOTA-Ala(SO3H)-Aminooctanoyl-Gln-Trp-Ala-Val-N methyl Gly-His-Statine-Leu-NH2

  • Lim, Jae Cheong;Choi, Sang Mu;Cho, Eun Ha;Kim, Jin Joo
    • Journal of Radiation Industry
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    • v.7 no.2_3
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    • pp.127-134
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    • 2013
  • Bombesin receptors are overexpressed in many kinds of human tumors. In particular, the gastrin releasing peptide receptor (GRPR) which is also called bombesin receptor subtype 2, has been identified in prostate cancer. In the present study, we developed a bombesin antagonist-based $^{177}Lu$-labeled peptide, $^{177}Lu$-DOTA-$Ala(SO_3H)$-Aminooctanoyl-Gln-Trp-Ala-Val-N methyl Gly-His-Statine-Leu-$NH_2$ (DOTA-sBBNA). DOTA-sBBNA was prepared using a solid phase synthesis method. It was labeled with $^{177}Lu$ by a high radiolabeling yield (>98%), and its Log P value was -2.05. The radiolabeled peptide was highly stable in serum incubation at $37^{\circ}C$ for 48 hr. A competitive displacement of $^{125}I-[Tyr^4]$-Bombesin on the PC-3 human prostate carcinoma cells revealed that the $IC_{50}$ value of the peptide was 6.76 nM indicating a highly nanomolar binding affinity for GRPR. These results suggest that $^{177}Lu$-DOTA-sBBNA can be a potential candidate for targeting prostate cancer, and further studies to evaluate its biological characteristics are needed.

Novel Bombesin Analogues Conjugated with DOTA-Ala(SO3H)-4 aminobenzoic acid and DOTA-Lys(glucose)-4 aminobenzoic acid: Synthesis, Radiolabeling, and Gastrin Releasing Peptide Receptor Binding Affinity

  • Lim, Jae Cheong;Choi, Sang Mu;Cho, Eun Ha;Kim, Jin Joo
    • Journal of Radiation Industry
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    • v.7 no.2_3
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    • pp.191-200
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    • 2013
  • In this study, a novel bombesin (BBN) analogues, DOTA-Ala($SO_3H$)-4 aminobenzoyl-Gln-Trp-Ala-Val-Gly-His-Leu-Met-$NH_2$ (DOTA-sBBN) and DOTA-Lys(glucose)-4 aminobenzoyl-Gln-Trp-Ala-Val-Gly-His-Leu-Met-$NH_2$ (DOTA-gluBBN), were synthesized and radiolabeled, and their binding affinities were evaluated. Peptides were prepared by a solid phase synthesis method and their purities were over 98%. DOTA is the chelating agent for $^{177}Lu$-labeling, and the DOTA-conjugated peptides were radiolabeled with $^{177}Lu$ by a high radiolabeling yield (>98%). The Log P values of DOTA-sBBN and DOTA-gluBBN were -2.20 and -2.79, respectively. 50.41% of $^{177}Lu$-DOTA-sBBN and 72.97% of $^{177}Lu$-DOTA-gluBBN were left undegraded by the serum incubation at $37^{\circ}C$ for 48 hr. A competitive displacement of $^{125}I-[Tyr^4]$-BBN on the PC-3 human prostate carcinoma cells revealed that 50% inhibitory concentration ($IC_{50}$) were 1.46 nM of DOTA-sBBN and 4.67 nM of DOTA-gluBBN indicating a highly nanomolar binding affinity for GRPR. Therefore, it is concluded that $^{177}Lu$-DOTA-sBBN and $^{177}Lu$-DOTA-gluBBN can be potential candidates as a targeting modality for the Gastrin-releasing peptide receptor (GRPR)-over-expressing tumors, and further studies to evaluate their biological and pharmacological characteristics are needed.

Distribution and Extraction Condition of Endoprotease and Exopeptidase from Viscera of Illex argentinus (원양산 오징어(Illex argentinus) 내장의 endoprotease 및 exopeptidases의 분포 및 추출조건 검토)

  • Kim, Hye-Suk;Heu, Min-Soo;Kim, Jin-Soo
    • Applied Biological Chemistry
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    • v.50 no.4
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    • pp.308-315
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    • 2007
  • For the effective use of squid processing by-products as food resources, the distribution and the extraction condition of endoprotease and exopeptidase from viscera of Illex argentinus were investigated. Crude protein and lipid contents of viscera of Illex argentinus were 17.2% and 16.9%, respectively. Regardless of kinds of extraction solution (water, 1% NaCl, 1% KCl and 1% NaCl- KCl) and extraction times (1-20 h), endoprotease activities from viscera of Illex argentinus on Hb, casein and azocasein (pH 6.0) were higher than those on casein and azocasein of the other pHs, thus indicating that the distribution of protein hydrolysing protease is distinctive in the weak acid pH range. Exopeptidase activities against LeuPNA and ArgPNA at pH 7.5 were relatively higher than endoprotease activity of the same pH. The results suggested that exopeptidase among proteases from viscera of Illex argentinus was reasonable for application in food industry compared to endoprotease. The activity in enzymes from viscera of Illex argentinus was the highest in the exopeptidase extracted with deionized distilled water at room temperature for 6-8 h. The optimal reaction conditions of crude enzyme from viscera of Illex argentinus were 7.5 for pH and $50-55^{\circ}C$ for temperature.

Moisturizing Effects and Composition Analysis of Proteoglycan Isolated from Chia (Salvia hispanica) Seed (치아 씨앗으로부터 분리한 단백다당체의 성분분석과 보습 효과)

  • Lee, Bum-Chun;Joo, Chul-Gue;Hur, Ji-Yeon;Lee, Keun-Ha;Kim, Young-Jin;Lee, Chan-Woo;Kim, Jin-Woong;Park, Yong-Il;Kim, Hyun-Sook;Choe, Tae-Boo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.36 no.2
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    • pp.121-128
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    • 2010
  • We investigated the effects of skin hydration and composition analysis of proteoglycan (chia seed polymer) produced from chia (Salvia hispanica) seed. The result showed that proteoglycan of chia seeds is composed of galactose (46.8 %), glucuronic acid (27.1 %), rhamnose (8.7 %), xylose (7.6 %), glucose (4.9 %), fructose (2.3 %), mannose (1.8 %), arabinose (0.9 %) and the amount of proteins contained is 31.3 mg/g with the constituent amino acid compositions (mg/g) of Asp (1.9), Glu (3.6), Ser (0.9), Gly (3.6), Thr (0.8), Arg (1.0), Ala (2.0), Tyr (0.4), Cys (4.8), Val (1.1), Phe (0.5), Ile (0.6), Leu (0.9). The molecular weight of the proteoglycan measured by GPC (Gel Permeation Chromatography) is the range of 100,000~250,000 Da and the average molecular weight is 170,000 Da. The moisturizing effects and trans-epidermal water loss (TEWL) of chia seed polymer in cosmetic products (O/W emulsion) were studied in vivo. Chia seed polymer showed good skin hydration effects when compared with sodium hyaluronate which is a common moisturizer. Taken all together, chia seed polymer should be a very useful cosmetic ingredient as a moisturizer and a protecting agent from various skin irritations.

An Action Mechanism of Substance P on the Tracheal Smooth Muscle Contraction in Rabbits (토끼 기관의 평활근 수축에 미치는 substance P 의 작용기전)

  • 명창률
    • Journal of Chest Surgery
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    • v.27 no.1
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    • pp.9-14
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    • 1994
  • Substance P[SP] has been known to be a peptide which may be plays a role as a neurotransmitter in central nervous system as well as peripheral autonomic nervous system. It has been reported that SP was widely distributed in the nerve of the tracheal smooth muscle and induced the muscle contraction. However, definite action mechanism of SP in the tracheal smooth muscle was not clear, yet. Thus, present experiment was performed to elucidate an effect of substance P and an action mechanism on contraction of the smooth muscle in rabbits. In order to find a neural mechanism to the effect of SP on the tracheal smooth muscle contraction, atropine sulfate, tetrodotoxin, propranol and phentolamine were administered at 10 min before the addition of SP. Otherwise,to find effect of SP antagonists on the action of SP, [D-Pro2, D-Try7,9]SP, [D-Arg1, D-Pro2, D-Trp7,9, Leu11]SP and [D-Pro4, D-Trp7,9]SP were administered as a same fashion. These following results were obtained. 1] SP induced contraction of the tracheal smooth muscle under resting condition and the contraction was increased dose-dependently. 2] Cholinergic blocker[atropine], neural blocker[tetrodotoxin] and adrenergic blocker[propranol and phentolamine] didn`t have an effect on the contractile response. 3] Three SP antagonists inhibited the contractile response. 4] Isoproterenol relaxed the contraction induced by SP. The above results suggested that SP induced contraction of the tracheal smooth muscle directly act to the smooth muscle in rabbits. The autonomic nervous system did not seem to participate in the SP action.

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Comparison of Quality, Physiochemical and Functional Property between Organic and Conventional Rice (유기쌀과 일반쌀의 품질, 이화학적 특성 및 기능성 비교)

  • Park, Jang-Hyun;Nam, Seung-Hee;Kim, Young-Ok;Kwon, Oh-Do;An, Kyu-Nam
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.5
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    • pp.725-730
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    • 2010
  • The differences between organic and conventional rice were studied from the point of view of quality, physiochemical property and functionality. It was concluded that organic rice showed a 6% lower complete rice ratio than conventional rice. However, organic rice exhibited more excellent physiochemical property on amylose or protein content, Toyo value, and acidity with 17.5%, 6.6%, 8.12%, and 8.7%, respectively. There was no difference between two kinds of rice on alkali digestion value and amino acids contents such as Asp, Ser, Glu, Ala, Leu, Arg. Among inorganic compounds of rice, four compounds including B, Mn, Fe, or Zn were slightly higher at organic rice with 1.1~2.7 mg/100 g, compared to conventional rice. Interestingly, total phenolic acid and phytic acid content were 89% or 23% higher at organic rice than conventional rice, respectively. MeOH extract from organic rice showed higher antioxidant activity with 26% than that of conventional rice with 22%. In contrast, conventional rice was favored than organic rice on stickness, taste, palatability fields at sensory evaluation.

NMR Studies of Metal-binding Luteinizing Hormone Releasing Hormone

  • Won, Ho-Shik
    • Bulletin of the Korean Chemical Society
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    • v.32 no.11
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    • pp.4021-4026
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    • 2011
  • Functions of the luteinizing hormone releasing hormone (LHRH) and its induced release by divalent metal ions have received great attention because this neurotransmitter subsequently regulates the secretion of luteinizing hormone (LH). Metal-LHRH complexes were synthesized by addition of various Cu(II),Ni(II),Zn(II) ions into LHRH in order to understand how the induced release of LHRH is possible. The degree of complexation was monitored by $^1H$, $^{13}C$-NMR chemical shifts, and final products were identified by Mass spectrometry. Solutionstate structure determination of Zn(II)-LHRH out of metal-complexes was accomplished by using NMR and NMR-based distance geometry (DG). Interproton distance information from nuclear Overhauser effect spectroscopy was utilized for structure determination. Structure obtained in this study has a cyclic conformation exhibiting a specific ${\alpha}$-helical turn with residue numbers His[2]-Leu[7] out of 10 amino acids. Comparison of chemical shifts and EPR studies of Ni(II),Cu(II)-LHRH complexes exhibit that these metal complexes have 4-coordination geometry.

Preparation of Pronase Hydrolysate from Alaska-pollack (명태단백 Pronase 가수분해물의 제조)

  • 서형주
    • The Korean Journal of Food And Nutrition
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    • v.8 no.4
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    • pp.335-343
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    • 1995
  • In order to enhance the utility of alaska-pollack, the optimum conditions for the preparation of pronase hydrolysate. The optimum temperature and pH for the hydrolysis of alaska-pollack by pronase were 4$0^{\circ}C$ and pH 7.0. The reaction time and enzyme concentration were 4 hr and 1,000 units per g of substrate. Under the above optimum conditions alaska-pollack was hydrolysed by pronase yielding a hydrolytic degree of about 89eye. The bitterness and hyrophobicity of pronase hydrolysate were decreased with increasing reaction time. Hydrophobic amino acids(Tyr, Met, Ala, flu, Leu, and Phe) were increased for 2 hr, but fur thor hydrolysis was showed decrease of hydrophobic amino acids content. Palatable amino acids (Asp, Glu, Pro, Ser, Thr and Gly) were increased with hydrolysis time.

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Leuconostoc mesenteroides CJNU 0147 and Lactobacillus casei CJNU 0588 Improve Growth of a Bifidobacterium lactis Strain in Co-cultures

  • Eom, Ji-Eun;Moon, Gi-Seong
    • Preventive Nutrition and Food Science
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    • v.16 no.4
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    • pp.386-389
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    • 2011
  • Previous studies have confirmed that fermented whey produced by Leuconostoc mesenteroides CJNU 0147 or Lactobacillus casei CJNU 0588 display bifidogenic growth stimulator (BGS) activity. The present study sought to determine if the strain itself can improve the growth of bifidobacteria in co-cultures. In reinforced clostridial medium (RCM), both strains stimulated the growth of a Bifidobacterium lactis strain during the exponential phase and also stimulated the growth during almost all growth phases in whey broth. Fermented whey containing viable Leu. mesenteroides CJNU 0147 and L. casei CJNU 0588 cells maintained viability of the B. lactis strain stored at $10^{\circ}C$ in MRS broth. Viable cell count of the B. lactis strain without the fermented whey was decreased to 5.6 log cfu/mL after 15 days, whereas that of the strain with the fermented whey was slightly increased to 7.1 log cfu/mL as compared with initial viable cell count of 6.9 log cfu/mL.