• Title/Summary/Keyword: proline

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Inhibition of Prolyl 4-Hydroxylase by Oxaproline Tetrapeptides In Vitro and Mass Analysis for the Enzymatic Reaction Products

  • Moon Hong-sik;Begley Tedhg P.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.1
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    • pp.61-64
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    • 2000
  • A series of 5-oxaproline peptide derivatives was synthesized and evaluated for its ability to inhibit the prolyl 4-hydroxylase in vitro. Structure-activity studies show that the 5-oxaproline sequences, prepared by the 1,3-dipolar cycloaddition of the C-methoxycarbonyl-N-mannosyl nitrone in the presence of the ethylene, are more active than the corresponding proline derivatives. Prolyl 4-hydroxylase belongs to a family of $Fe^{2+}-dependent$ dioxygenase, which catalyzes the formation of 4-hydroxyproline in collagens by the hydroxylation of proline residues in -Gly-Xaa-Pro-Gly- of procollagen chains. In this paper we discover the more selective N-Cbz-Gly-Phe-Pro-Gly-OEt $(K_m\;=\;520\;{\mu}M)$ sequences which are showed stronger binding than others in vitro. Therefore, we set out to investigate constrained tetrapeptide that was designed to mimic the proline structure of pep tides for the development of prolyl 4-hydroxylase inhibitor. From this result, we found that the most potent inhibitor is N-Dansyl-Gly-Phe-5-oxaPro-Gly-OEt $(K_i\;=\;1.6\;{\mu}M)$. This has prompted attempts to develop drugs which inhibit collagen synthesis. Prolyl 4-hydroxylase would seem a particularly suitable target for antifibrotic therapy.

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Novel p104 protein regulates cell proliferation through PI3K inhibition and p27Kip1 expression

  • Han, Seung-Jin;Lee, Jung-Hyun;Choi, Ki-Young;Hong, Seung-Hwan
    • BMB Reports
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    • v.43 no.3
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    • pp.199-204
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    • 2010
  • The protein p104 was first isolated as a binding partner of the Src homology domain of phospholipase C$\gamma$1, and has been shown to associate with p85$\alpha$, Grb2. The ectopic expression of p104 reduced cellular growth rate, which was also achieved with the overexpression of only the proline-rich region of p104. The proline-rich region of p104 has been found to inhibit the colony formation of platelet-derived growth factor BB-stimulated NIH3T3 cells and MCF7 cancer cells on soft agar. Mutagenesis analysis showed that the second and third proline-rich regions are essential for growth control, as well as for interaction with p85$\alpha$. Overexpression of p104 increased the level of the cyclin-dependent kinase inhibitor, $p27^{Kip1}$, and inhibited the activity of phosphoinositide 3-kinase (PI3K). In summary, p104 interacts with p85$\alpha$ and is involved in the regulation of $p27^{Kip1}$ expression for the reduction of cellular proliferation.

Conformational Preference of Pseudo-Proline Dipeptide in the Gas Phase and Solutions

  • Park, Hae-Sook;Kang, Young-Kee
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.74-74
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    • 2003
  • We report here the results on N-acetyl-N'-methylamide of oxazolidine (Ac-Oxa-NHMe) calculated using the ab initio molecular orbital method with the self-consistent reaction field (SCRF) theory at the HF level of theory with the 6-3l+G(d) basis set. The displacement of the $\square$-CH$_2$ group in proline ring by oxygen atom has affected the structure of proline, cis$\^$∼/ trans equilibrium, and rotational barrier. The up-puckered structure is found to be prevalent for the trans conformers of the Oxa amide. The higher cis populations of the Oxa amide can be interpreted due to the longer distance between the acetyl methyl group and the 5-methylene group of the ring for the trans conformer of the Oxa amide than that of the Pro amide. The changes in charge of the prolyl nitrogen and the decrease in electron overlap of the C$\^$∼/ N bond for TS structures seem to play a role in lowering rotational barriers of the Oxa amide compared to that of the Pro amide. The calculated preferences for cis conformers in the order of Oxa > Pro amides and for trans-to-cis rotational barriers in the order of Pro > Oxa amide in water are consistent with experimental results on Oxa-containing peptides. The pertinent distance between the prolyl nitrogen and the N$\^$∼/ H amide group to form a hydrogen bond might indicate that this intramolecular hydrogen bond could contribute in stabilizing the TS structures of Oxa and Pro amides and play a role in prolyl isomerization.

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Inhibition of Adventitious Root Growth in Boron-Deficient or Aluminum-Stressed Sunflower Cuttings

  • Hong, Jung-Hee;Go, Eun-Jung;Kim, Tae-Yun
    • Journal of Environmental Science International
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    • v.12 no.11
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    • pp.1189-1196
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    • 2003
  • The effect of boron and aluminum on the development of adventitious roots was studied in sunflower cuttings. Three-day-old seedlings were de-rooted and grown in nutrient solutions with or without boron and supplemented with different concentrations (from 50 to 700 ${\mu}$M) of aluminum. The number and length of the adventitious roots and proline content in adventitious roots in response to insufficient boron and aluminum stress were determined periodically. The micronutrient boron caused the development of numerous roots in the lower parts of the hypocotyl. A dose-response of boron-induced rooting yielded an optimum concentration of 0.1 mM boron. In the absence of boron, in the majority of the adventitious roots, a significant inhibition was observed with or without aluminum, indicating that the most apparent symptom of boron deficiency is the cessation of root growth. Increasing concentrations of aluminum caused progressive inhibition of growth and rooting of the hypocotyls, and a parallel increase in proline levels of adventitious roots. Supplemental boron ameliorated the inhibitory effect of aluminum, suggesting that aluminum could inhibit root growth by inducing boron deficiency. Ascorbate added to medium in the absence of boron improved root growth and induced a significant decrease in proline levels. These findings suggest that adventitious root growth inhibition resulting from either boron deficiency or aluminum toxicity may be a result of impaired ascorbate metabolism.

Analysis of Mahjor Flavor Compounds in Takju Mash Brewed with a Modified Nuruk (개량누룩으로 제조한 탁주의 중요 향미성분 분석)

  • 이영숙
    • The Korean Journal of Food And Nutrition
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    • v.12 no.4
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    • pp.421-426
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    • 1999
  • The major flavor compounds of Takju mash which was brewed with a modified Nuruk made by inocu-lation and cultivation of Rhizopus japonicus T2, Aspergillus oryzae L2 and Hansenula sp. BC26 isolated from Nuruk, were analyzed, as compared with those with current fermenting agents such as commerical Nur-uk and rice koji of Aspergillus kawachii. The contents of isoamyl alcohol isobutyl alcohol and ethyl acet-ate which were known as aroma compounds in Takju were much higher in mash of modified Nuruk than in that of commercial Nuruk or ricd koji. The major organic acids were lactic fumalic and succinic acid in mash of modified and lactic and acetic acid in mash of commercial Nuruk and citric lactic and suc-cinic acid in mash of rice koji. The contents of total organic acids were 5,146mg/L, 1,706mg/L and 1, 388 mg/L in mash of commercial Nuruk rice koji and modified Nuruk respectively. The major free amino acids were glutamic acid alanine proline and histidine in mash of modified Nuruk and glutamic acid proline leucine and histidine in mash of commercial Nuruk and arginine proline and glutamic acid in mash of rice koji. The contents of total free amino acids were 14,090mg/L 12,202mg/L and 7,152 mg/L in mash of modified Nuruk commercial Nurcuk and rice koji respectively. Therefore it seemed that the Takju mash of modified Nuruk was better than that of commercial Nuruk or rice koji.

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Salt tolerance and proline accumulation of potato (Solanum tuberosum L.) in vitro plants to NaCl treatment (NaCl 처리에 따른 감자 (Solanum tuberosum L.) 기내 식물체의 내염성 및 Proline 함량 변화)

  • Im, Ju Sung;Cho, Ji Hong;Cho, Kwang Soo;Chang, Dong Chil;Jin, Yong Ik;Yu, Hong Seob;Cheun, Chung Gi;Kim, Mee Ok;Han, Da Som;Lee, Seul Ki;Kim, Wha Yeong
    • Journal of Plant Biotechnology
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    • v.42 no.2
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    • pp.129-134
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    • 2015
  • In order to compare salt tolerance among potato cultivars (Solanum tuberosum L.), in vitro plants of each cultivar were cultured on the Murashige and Skoog's medium containing different levels of NaCl (0, 75, 150, and 225 mM). The suitable level of NaCl for a comparison of the shoot and root growth between potato cultivars was 75mM. Shoot length and weight were better in 'Dejima', 'Superior', 'Jayoung', and 'Haryeong' than in other cultivars such as 'Goun' and 'Atlantic'. Normal root growth was observed in 'Seohong', 'Superior', and 'Haryeong', while 'Goun', 'Atlantic', 'Dejima', 'Jowon', and 'Chuback' showed no root. Proline contents in all the cultivars increased as affected by NaCl levels, then the increase amount was fewer in the cultivars showed a better growth in both shoot and root than in others showed a poor growth. As a result of this study, 'Superior' and 'Haryeong' were determined as salt tolerant cultivars and therefore using these cultivars it would be possible to potato production at salt arable land such as reclaimed tidal.

Characterization of the Acetolactate synthase (ALS) gene and Molecular Assay of Mutations Associated with Sulfonylurea Herbicide Resistance of Monochoria vaginalis (물달개비의 Acetolactate synthase (ALS) 유전자의 특성과 Sulfonylurea 제초제 저항성과 관련 돌연변의 분자생물학적 접근)

  • Park, Tae-Seon;Park, Hong-Kyu;Ku, Bon-Il;Kim, Young-Doo;Ko, Jae-Kwon;Lee, In-Yong;Park, Jae-Eup
    • The Korean Journal of Pesticide Science
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    • v.13 no.4
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    • pp.290-297
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    • 2009
  • This research aims to contribute the characterization of acetolactate synthase (Ec 4.1.3.18; ALS) and the resistance mechanism by sequence analysis of ALS gene of the sulfonylurea-resistant and -susceptible Monochoria vaginalis. The ALS gene was obtained from susceptible (S) and resistant (R) M. vaginalis to sulfonylurea herbicides (SUs). The 815 bp the fragment and the genomic DNA sequence coding for acetolactate synthase (ALS) of S and R biotypes of M. vaginalis were cloned and sequenced. Nineteen clones were divided greatly into 4 groups as result of sequencing. The first group was not difference to S type, the second group was amino acid of P197S which found point mutations causing substitution of serine for proline at amino acid 197, the third group was observed greatly other part of 6 places than group 1, and the fourth group appeared the intergrade of group 1 and 3. Therefore, it could be assumed what ALS gene of various types can be one plant. The peptide of the 13 amino acid Domain A region for ALS genes from R biotype of M. vaginalis differed from that of the S biotype by one base substitution at proline codon of Domain A. It could also be confirmed that point mutation of serine for proline at amino acid 197.

Effects of Mycorrhizal Inoculation on Plant Growth and N Metabolites in Relation to drought-stress Tolerance (Mycorrhiza 접종이 가뭄 스트레스하의 식물성장과 질소 대사산물에 미치는 영향)

  • Lee, Bok-Rye;Jung, Woo-Jin;Kim, Dae-Hyun;Kim, Kil-Yong;Shon, Bo-Kyoon;Kim, Tae-Hwan
    • Korean Journal of Soil Science and Fertilizer
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    • v.35 no.5
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    • pp.314-325
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    • 2002
  • The effects of arbuscular mycorrhizal (AM) fungus (Glomus intraradices) on plant growth and N metabolic responses were examined in perennial ryegrass plants exposed to drought-stressed or well-watered condition. Mycorrhizal inoculation improved significantly leaf water potential, dry mass and P content. Drought stress increased significantly nitrate concentration in roots where the increase was much less in AM than non-AM. Drought stress decreased the concentration of soluble proteins in non-AM shoots, whereas non-significant decline occurred in AM shoots even under drought condition. The concentrations of ammonia and proline in drought stressed non-AM plants significantly increased, while mycorrhizal inoculation lowered significantly ammonia and proline accumulation. The decrease in leaf dry weight in drought stressed-plants was significantly correlated to the increase in ammonia (p<0.01) and proline concentration (p<0.01). These results suggested that the increased P content and N assimilation by mycorrhizal inoculation may be associated with drought stress tolerance, showing the moderating effects on shoot growth inhibition and ammonia accumulation in drought stressed-plants.

Free Amino Acid and Nitrogen Contents of the Coastal Plants in Korea (해안식물의 유리아미노산과 질소함량)

  • 추연식;도정화;송승달
    • The Korean Journal of Ecology
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    • v.22 no.3
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    • pp.109-117
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    • 1999
  • Thirty two species of coastal plants (mainly halophytes) were investigated for their free amino acids and the total and water-soluble nitrogen contents in leaves. All plants except some species (e.g. Scutellaria and Linaria) contained proline, but only Ageratum. Tetragonia and Raphanus in rather high amounts, that it can be thought to serve as a cytoplasmic osmolyte. In some plant species (Euphorbia. Glehnia. Peucedanum. Raphanus and monocotyledonous Carex and Zoysia), however, hydroxyproline. (OH-Pro) rather than proline were accumulated to a considerable extent. The concentrations of total free amino acids were low in Aster tripolium, Linaria. Lysimachia. Plantago. Rumex, Vitex and especially in the members of the Chenopodiaceae and Crassulaceae. Marked differences also occurred in the nitrogen levels. Aizoaceae, Chenopodiaceae, Convolvulaceae, Cruciferae, Euphorbiaceae and Leguminosae usually showed high values of total and soluble amino nitrogen, while the opposite was true for most of the Cyperaceae, Gramineae, Lamiaceae, Plantaginaceae, Scophuriaceae, and Verbenaceae. The free amino acids in the investigated plant species contributed very little to the nitrogen content, but in plants of Euphorbia, Messerschmidia and Orostachys. their amino acid-N made up for 25∼30% of the total nitrogen. In conclusion, only a few cases did proline known as compatible solute constitute a significant proportion of the free amino acid pool in coastal plants.

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Efficient Callus Induction and Plant Regeneration of Guineagrass (Panicum maximum Jacq.) (난지형 목초 기니아그라스의 효율적인 캘러스 유도 및 식물체 재분화)

  • Seo, Mi-Suk;Takahara, Manabu;Takamizo, Tadashi
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.30 no.4
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    • pp.283-290
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    • 2010
  • Guineagrass (Panicum maximum Jacq.) is an important warm-season forage grass as well as biomass crop. It has both sexual and asexual mode of reproduction (apomictic) depending on cultivar. We developed efficient plant regeneration system for an apomictic (cv. Natsukaze) and a non-apomictic (Noh-PL1) guineagrass by optimizing the level of L-proline in the callus induction and that of $AgNO_3$ in plant regeneration medium. Among the L-proline concentrations tested, the best callus induction was achieved by using 2g/L L-proline in both the genotypes. Immature embryos proved to be the best explant source for tissue culture of guineagrass. The highest frequency of shoot regeneration was obtained on MS plant regeneration medium supplemented with 2 mg/L $AgNO_3$. These results provide a foundation for efficient tissue culture and genetic improvement of guineagrass.