• Title/Summary/Keyword: amino acid inhibition

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EXPRESSION AND FUNCTIONAL CHARACTERIZATION OF AMINO ACID TRANSPORT SYSTEM L IN SAOS2 HUMAN OSTEOGENIC SARCOMA CELLS (사람 골육종 세포 Saos2에서 아미노산 수송계 L의 발현 및 기능적 특성)

  • Kim, Su-Gwan;Kim, Hyun-Ho;Kim, Chang-Hyun;Kim, Do-Kyung
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.32 no.3
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    • pp.200-208
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    • 2006
  • Amino acids are required for protein synthesis and energy sources in all living cells. The amino acid transport system L is a major nutrient transport system that is responsible for $Na^+$-independent transport of neutral amino acids including several essential amino acids. In malignant tumors, the L-type amino acid transporter 1 (LAT1), the first isoform of system L, is highly expressed to support tumor cell growth. In the present study, the expression and functional characterization of amino acid transport system L were, therefore, investigated in Saos2 human osteogenic sarcoma cells. RT-PCR and western blot analyses have revealed that the Saos2 cells expressed the LAT1 and the L-type amino acid transporter 2 (LAT2), the second isoform of system L, together with their associating protein heavy chain of 4F2 antigen (4F2hc) in the plasma membrane, but the expression of LAT2 was very weak. The uptakes of [${14}^C$]L-leucine by Saos2 cells were $Na^+$-independent and were completely inhibited by the system L selective inhibitor, 2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid (BCH). The affinity of [${14}^C$]L-leucine uptake and the inhibition profiles of [${14}^C$]L-leucine uptake by various amino acids in the Saos2 cells were comparable with those for the LAT1 expressed in Xenopus oocytes. The majority of [${14}^C$]L-leucine uptake is, therefore, mediated by LAT1 in the Saos2 cells. These results suggest that the transports of neutral amino acids including several essential amino acids into Saos2 human osteogenic sarcoma cells are for the most part mediated by LAT1. Therefore, the Saos2 human osteogenic sarcoma cells are excellent tools for examine the properties of LAT1. Moreover, the specific inhibition of LAT1 in tumor cells might be a new rationale for anti-tumor therapy.

Leaf Exudates of Vicia faba and their Effects on Botrytis fabae and Some Associated Fungi

  • Migahed, Fatma F.;Nofel, Ashraf M.
    • Mycobiology
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    • v.29 no.4
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    • pp.198-204
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    • 2001
  • Analysis of leaf exudates of Vicia faba using paper chromatography to identify individual amino acids and sugars qualitatively was investigated. The results revealed that the number of identified amino acids detected in the leaf exudates of the susceptible plants was more than those of resistant plants. The results also showed an increase in the number of amino acids exuded by infected leaves, but no marked difference in sugars of infected and non infected plants. Lithium chloride application led to decrease in amino acid and sugar contents. The number of amino acids and sugars was also decreased with leaf age. Botrytis fabae and the selected fungal species(Alternaria alternata, Fusarium oxysporum and Aspergillus niger) were used to show the effect of individual amino acid and sugar on their spore germination. It was observed that all amino acids stimulated the fungal spore germination except serine which inhibited its spore germination. In case of A. alternata, spore germination was stimulated by all amino acids except serine, alanine, glutamic acid, arginine and methionine which caused the inhibition. In case of F. oxysporum, aspartic and glutamic acids inhibited spore germination but the other amino acids stimulated its spore germination. Aspartic acid and phenyl alanine inhibited the spore germination of A. niger. All the identified sugars(galactose, glucose, fructose and rhamnose) stimulated spore germination of all tested fungi.

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Comparison of the Nucleotide Sequence of Cloned Osteopontin from Hanwoo and Holstein

  • Lee, Tae Young;Ju, Sung Kyu;Nam, Myoung Soo
    • Food Science of Animal Resources
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    • v.33 no.3
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    • pp.331-334
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    • 2013
  • Osteopontin (OPN) is a secreted phosphorylated glycoprotein. It has an important role in multiple biological processes including cell survival, bone remodeling, inhibition of ectopic calcification, as well as, is thought to have potential immune modulation activities. In this work, we isolated and characterized a full-length open reading frame (ORF) of Korean native cow's OPN from Korean native cow's (Hanwoo) kidney, and successfully cloned firstly on Hanwoo's OPN. The sequencing results indicated that the isolated cDNA was 1190 bp in length containing a complete ORF of 837 bp. It encoded a precursor protein Hanwoo's OPN consisting of 278 amino acids with a signal peptide of 16 amino acids. Amino acid homology was found to be 99.3% as compared to the corresponding sequences of Holstein bone marrow OPN. Hanwoo's kidney OPN and Holstein bone marrow OPN are different only in two amino acid residues 42 and 56, amino acid residue 42 is Thr (T) ${\leftrightarrow}$ Ile (I), and amino acid residue 56 is Ala (A) ${\leftrightarrow}$ Thr (T) respectively. These results from the present work would be helpful to elucidate the biological function of Hanwoo's OPN and provided a foundation for further insight into role of Hanwoo's OPN.

Mechanism of Growth Inhibition by BCH in HEp2 Human Head and Neck Squamous Cell Carcinoma (사람 두경부 편평세포암종 HEp2 세포에서 BCH에 의한 세포성장 억제기전)

  • Choi, Bong-Kyu;Jung, Kyu-Yong;Cho, Seon-Ho;Kim, Chun-Sung;Kim, Do-Kyung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.5
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    • pp.555-560
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    • 2008
  • Amino acid transporters are essential for the growth and proliferation in all living cells. Among the amino acid transporters, the system L amino acid transporters are the major nutrient transport system responsible for the $Na^+$-independent transport of neutral amino acids including several essential amino acids. The L-type amino acid transporter 1 (LAT1), an isoform of system L amino acid transporter, is highly expressed in cancer cells to support their continuous growth and proliferation. 2-Aminobicyclo-(2,2,1)-heptane-2-carboxylic acid (BCH) is a model compound for the study of amino acid transporter as a system L selective inhibitor. We have examined the effect and mechanism of BCH on cell growth suppression in HEp2 human head and neck squamous cell carcinoma. The BCH inhibited the L-leucine transport in a concentration-dependent manner with a $IC_{50}$ value of $51.2{\pm}3.8{\mu}M$ in HEp2 cells. The growth of HEp2 cells was inhibited by BCH in the timeand concentration-dependent manners. The formation of DNA ladder was not observed with BCH treatment in the cells. Furthermore, the proteolytic processing of caspase-3 and caspase-7 in the cells were not detected by BCH treatment. These results suggest that the BCH inhibits the growth of HEp2 human head and neck squamous cell carcinoma through the intracellular depletion of neutral amino acids for cell growth without apoptotic processing.

Developement of novel enzyme system for production of enantiomerically pure ${\beta}-amino$ acids : Kinetic resolution of racemic 3-amino-n-butanoic acid using transaminase from Alcaligenes denitrificans Y2k-2

  • Im, Seong-Yeop;Jo, Byeong-Gwan;Kim, Byeong-Gi
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.579-582
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    • 2000
  • (R,S)-3-amino-n-butanoic acid$(DL-\;{\beta}\;-homoalanine)$ has been kinetically resolved using Alcaligenes denitrificans Y2k-2 as a biocatalyst, which was isolated from soil by enrichment culture, which was carried out with minimal media containing (R,S)-3-amino-n-butanoic acid as a sole nitrogen source. The enzyme which peformed this kinetic resolution assumed to belong to the ${\omega}-transaminase$ family, because A. denitrificans used pyruvate as amino acceptor and its transaminase activity was inhibited by gabaculine, aminooxy acetic acid and hydroxylamine. In whole cell reaction, (R,S)-3-amino-n-butanoic acid was kinetically resolved to the corresponding (R)-3-amino-n-butanoic acid with excellent E (>100) in the presence of pyruvate as an amino acceptor at $37^{\circ}C$. (S-specific) We observed the substrate inhibition for pyruvate at 100mM. In this study, characteristics of transaminase activity of Alcaligenes denitrificans Y2k-2, such as substrate specificity and thermostability, are carried out for the development of (R)-3- amino-n-butanoic acid production system.

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Prolyl Endopeptidase Inhibitory Activity of 6-O-Palmitoyl L-Ascorbic Acid

  • Park, Yoon-Seok;Paik, Young-Sook
    • Journal of Applied Biological Chemistry
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    • v.49 no.3
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    • pp.110-113
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    • 2006
  • Prolyl endopeptidase (PEP, EC 3.4.21.26, also referred to as prolyl oligopeptidase) degrades proline containing, biologically active neuropeptides such as vasopressin, substance P and thyrotropin-releasing hormone by cleaving peptide bonds on carboxyl side of prolyl residue within neuropeptides of less than 30 amino acids. Evaluation of PEP levels in postmortem brains of Alzheimer's disease patients revealed significant increases in PEP activity. Therefore, a specific PEP inhibitor can be a good candidate of drug against memory loss. Upon our examination for PEP inhibitory activity from micronutrients, ascorbic acid (vitamin C) showed small but significant PEP inhibition (13% PEP inhibition at $8{\mu}g{\cdot}ml^{-1}$). Palmitic acid showed almost no PEP inhibition. However, 6-O-palmitoyl ascorbic acid ($\underline{1}$) showed 70% PEP inhibition at $8{\mu}g{\cdot}ml^{-1}$ indicating that hydrophobic portion of the compound $\underline{1}$ may facilitate the inhibitory effect. $IC_{50}$ value of compound $\underline{1}$ was $12.6{\pm}0.2{\mu}M$. The primary and secondary Lineweaver Burk and Dixon plots for compound $\underline{1}$ indicated that it is a non-competitive inhibitor with inhibition constant (Ki) value of $23.7{\mu}M$.

The Browning Reaction and Inhibition of Apple Concentrated Juice (사과농축액의 갈변현상 및 그 억제)

  • 배수경;이영철;김현위
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.1
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    • pp.6-13
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    • 2001
  • The study was conducted to investigate the effect of the browning inhibitors such as PVPP(polyvinylpoly-pyrrolidone), A.A.(ascorbic acid) on nonezymatic browning factors [free sugar, total amino acid, organic acid, A.A., HMF (hydroxymethylfurfural)] and enzymatic browning factors [PRO (polyphenoloxidase) activity, polyphenol compounds] in concentrated apple juice during 90 days storage. Considering color value (L value, $\Delta$E), absorbance at 420 nm, concentrated apple juice during 90 days storage. Considering color the effect of browning inhibition. According to the storage period, the changes of nonenzymatic factors in concentrated apple juice added with browning inhibitors were similar to those in control (concentrated apple juice without browning inhibitors), which were the decreased of sucrose(0.24~0.35% at 90 days), the slight increase of glucose and fructose, the decrease of total amino acid (530.4~573.1 mg/10g at 90 days), same value of A.A. at 90 days (38.5~78.6 mg/100g), and the increase of HMF (27.8~30.6 mg/100g at 90 days). On the contrary, enzymatic browning factors were significantly inhibited in concentrated apple juice added with PVPP, judging from the slow increase of PRO activity and the significant decrease of initial value in polyphenol compounds (especially chlorogenic acid). These results suggest that PVPP plays an important role as enzymatic browning inhibitor, that is, a scavenger of polyphenol compounds by adsorption in concentrated apple juice.

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Effects of Amino Acids on the Inhibition of Polyphenol Oxidase Activity from Perillae Folium (아미노산류가 들깨잎 폴리페놀 옥시다제 활성저해에 미치는 영향)

  • Park, Soo-Sun;Kim, An-Keun;Sohn, Eun-Soo
    • YAKHAK HOEJI
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    • v.40 no.1
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    • pp.65-71
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    • 1996
  • Characterization of Polyphenol oxidase (PPO) in Perillae Folium, particullarly inhibitor studies were investigated. This enzyme was stable at pH 5.0 and the residual activity of PPO at ${\geq}$ ph 5.5 was estimated to be very low. PPO activity was decreased slightly by adding amino acid with catechol as a substrate, particullary PPO activity was inhibited markedly by cystein, histidine, lysine and arginine. In the absorption spectra of the product formed when catechol was oxidized by PPO, with a ${\lambda}_{max}$ at 410nm, the peak shifted toward ${\lambda}_{max}$ 520nm by addition of L-proline. At relatively low concentrations($10^{-3}M$), sulfite and dithiothreithol completely inhibited PPO activity. Inhibition of PPO activity by amino acids and inhibitors increased or decreased depending on the pH used to measure it.

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Antioxidant and Antihypertensive Activities of Grains Grown in South Korea in Relation to Phenolic Compound and Amino Acid Contents

  • Narae Han;Koan Sik Woo;Jin Young Lee;Jiho Chu;Mihyang Kim;Yu-Young Lee;Moon Seok Kang;Hyun-Joo Kim
    • The Korean Journal of Food And Nutrition
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    • v.36 no.6
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    • pp.572-580
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    • 2023
  • Hypertension is characterized by excessive renin-angiotensin system activity, leading to blood vessel constriction. Several synthetic compounds have been developed to inhibit renin and angiotensin-converting enzyme (ACE). These drugs often have adverse side effects, driving the exploration of plant protein-derived peptides as alternative or supplementary treatments. This study assessed the phenolic compound and amino acid content and the antioxidant and antihypertensive activity of 5 South Korean staple crops. Sorghum had the highest phenolic compound content and exhibited the highest antioxidant activity. Millet grains, particularly finger millet (38.86%), showed higher antihypertensive activity than red beans (14.42%) and sorghum (17.16%). Finger millet was found to contain a large proportion of branched-chain, aromatic, and sulfur-containing amino acids, which are associated with ACE inhibition. In particular, cysteine content was positively correlated with ACE inhibition in the crops tested (r=0.696, p<0.01). This study confirmed that the amino acid composition was more correlated with the antihypertensive activity of grains than the phenolic compound content. Finger millet mainly contained amino acids, which have higher ACE inhibitory activity, resulting in the strongest antihypertensive activity. These findings underscore the antihypertensive potential of select crops as plant-based food ingredients, offering insight into their biological functions.

Analysis of the Change of Amino Acids by Abscisic acid and Polyamine Treatment in Radish Young Cotyledons (무우 유식물의 자엽에서 Abscisic acid와 Polyamine 처리에 의한 아미노산 변화의 분석)

  • Cho, Bong-Heuy
    • Analytical Science and Technology
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    • v.12 no.5
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    • pp.441-446
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    • 1999
  • The changes of amino acids in the cotyledons of spring - and winter radishes were analysed during the abscisic acid (ABA). ABA plus cycloheximide (CH) and ABA plus polyamine (PA) treatment. The total contents of amino acids were increased in spring radishes, and decreased in winter radishes by ABA. The contents of hydrophilic amino acids, proline, glycine, serine and cysteine in spring radishes (expecially proline), and of cysteine, leucine and phenylalanine in winter radishes (expecially cysteine and leucine)were increased, CH treatment resulted in the accumulation of amino acids by the inhibition of new synthesized protein, which synthesized against ABA induced dehydration both of radishes. On the contrary, the contents of amino acids were decreased in spring radishes, and increased in winter radishes during PA treatment. These results indicated that the accumulated hydrophilic amino acids and new synthesized proteins induced the adaptation of dehydration against stress and the role of ABA in accelerating of stress adaptation was mediated by polyamines.

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