• Title/Summary/Keyword: asparagine

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Effects of Oral Taurine Supplementation on Plasma Concentration and Urinary Excretion of Free Amino Acids in Healthy Female Adults (타우린복용이 정상 성인여성의 혈장 유리아미노산 농도 및 소변내 배설에 미치는 영향)

  • 차희숙
    • Journal of Nutrition and Health
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    • v.32 no.2
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    • pp.158-165
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    • 1999
  • Effects of oral taurine supplementation (6g/day) on plasma concentration and urinary of free amino acids were evaluated in healthy female adults. Among twenty five female volunteers(23.6$\pm$0.3 years old) participated in the taurine supplementation program, twenty four subjects successfully completed the two supplementation program. Plasma and urinary levels of free amino acids were determined by using an automated amino acid analyzer based on ion-exchange chromatography. Two weeks of taurine supplementation resulted in a 65% increase in plasma taurine concentration (p<0.001), Changes in fasting plasma amino acid concentrations followed by taurine supplementation were not spectacular, and were all within the normal range for human aldults. Taurine supplementation significantly elevated urinary methionine, asparagine, hydorxyproline and phosphoserine excretions(31~280%), and significantly decreased the urinary excretions of isoleucine, glutamate and serine compared to the values prior to taurine supplementation. For almost every individual amino acids, 24 hr urinary excretion level was significantly correlated to the urinary excretion value expressed as nmol/mg creatinine(p<0.001). A significant negative correlation found between plasma glutamine concentration and urinary glutamine excretion level suggests that the decrease in plasma glutamine concentration might be associated with the enhanced glutamine excretion in urine followed by taurine supplementation.

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Optimum Culture Conditions for ${\alpha}-Amylase$ Inhibitor Production of Streptomyces minoensis DMCJ-144, ${\alpha}-Amylase$ Inhibitor Producing Actinomycetes (${\alpha}$-아밀라아제 저해제 생성 방선균, Streptomyces minoensis DMCJ-144의 저해제 생산을 위한 최적 배양 조건)

  • Seo, Seong-Ok;Choi, Eung-Chil;Kim, Byong-Kak
    • YAKHAK HOEJI
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    • v.36 no.4
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    • pp.390-396
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    • 1992
  • Streptomyces minoensis DMCJ-144 isolated from soil produces the ${\alpha}-amylase$ inhibitor. Optimum culture conditions for ${\alpha}-amylase$ inhibitor production of the strain were determined in this experiment. The optimum composition of the culture medium was studied by supplementing various carbon sources, nitrogen sources, vitamins, and metal salts to the basal medium containing 1% glucose, 0.1% asparagine, 0.005% $MgSO_4{\cdot}7H_2O$, 0.005% $K_2HPO_4$, 0.005% NaCl. Other culture conditions such as the culture temperature, initial pH of the medium, aeration, and culture time were also investigated. When the strain was cultured in a 100 ml flask containing 20 ml of 2% glucose, 0.5% beef extract, 0.0002% riboflavin, 0.0002% thiamine HCI, 0.01% $ZnCl_2$, 0.005% $MgSO_4{\cdot}7H_2O$, 0.005% $CuSO_4{\cdot}5H_2O$, 0.005% NaCl, pH 7.2, 180 rpm at $30^{\circ}C$, the maximum production of the ${\alpha}-amylase$ inhibitor was observed after 5 days of the cultivation.

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Irreversible Thermoinactivation Mechanisms of Subtilisin Carlsberg

  • Dong Uk Kim
    • Bulletin of the Korean Chemical Society
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    • v.10 no.6
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    • pp.600-604
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    • 1989
  • In order to find the rational methods for improving the thermal stability of subtilisin Carlsberg, the mechanisms of irreversible thermoinactivation of the enzyme were studied at $90^{\circ}C.$ At pH 4, the main process was hydrolysis of peptide bond. This process followed first order kinetics, yielding a rate constant of $1.26\;{\times}\;10^{-1}h^{-1}$. Hydrolysis of peptide bond of PMS-subtilisin occurred at various sites, which produced new distinct fragments of molecular weights of 27.2 KD, 25.9 KD, 25.0 KD, 22.3 KD, 19.0 KD, 17.6 KD, 16.5 KD, 15.7 KD, 15.0 KD, 13.7 KD, and 12.7 KD. Most of the new fragments originated from the acidic hydrolysis at the C-side of aspartic acid residues. However 25.0 KD, 15.7 KD, and 13.7 KD which could not be removed in purification steps stemmed from the autolytic cleavage of subtilisin. The minor process at pH 4 was deamidation at asparagine and/or glutamine residues and some extend of aggregation was also observed. However, the aggregation was main process at pH 7 with a first order kinetic constant of $16 h^{-1}.$ At pH 9, the main process seemed to be combination of deamidation and cleavage of peptide bond.

Metabolite analysis in the type 1 diabetic mouse model

  • Park, Sung Jean
    • Journal of the Korean Magnetic Resonance Society
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    • v.25 no.3
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    • pp.33-38
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    • 2021
  • Type 1 diabetes mellitus (T1DM) is caused by insufficient production of insulin, which is involved in carbohydrate metabolism. Type 2 diabetes mellitus (T2DM) has insulin resistance in which cells do not respond adequately to insulin. The purpose of this study was to estimate the characteristics of type 1 diabetes using streptozotocin-treated mice (STZ-mouse). The sera samples were collected from the models of hyperglycemic mouse and healthy mouse. Based on the pair-wise comparison, five metabolites were found to be noticeable: glucose, malonic acid, 3-hyroxybutyrate, methanol, and tryptophan. It was very natural glucose was upregulated in STZ-mouse. 3-hyroxybutyrate was also increased in the model. However, malonic acid, tryptophan, and methanol was downregulated in STZ-mouse. Several metabolites acetoacetate, acetone, alanine, arginine, asparagine, histidine, lysine, malate, methionine, ornithine, proline, propylene glycol, threonine, tyrosine, and urea tended to be varied in STZ-mouse while the statistical significance was not stratified for the variation. The multivariate model of PCA clearly showed the group separation between healthy control and STZ-mouse. The most significant metabolites that contributed the group separation included glucose, citrate, ascorbate, and lactate. Lactate did not show the statistical significance of change in t-test while it tends to down-regulated both in DNP and Diabetes.

Genetic Background Behind the Amino Acid Profiles of Fermented Soybeans Produced by Four Bacillus spp.

  • Jang, Mihyun;Jeong, Do-Won;Heo, Ganghun;Kong, Haram;Kim, Cheong-Tae;Lee, Jong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.31 no.3
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    • pp.447-455
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    • 2021
  • Strains of four Bacillus spp. were respectively inoculated into sterilized soybeans and the free amino acid profiles of the resulting cultures were analyzed to discern their metabolic traits. After 30 days of culture, B. licheniformis showed the highest production of serine, threonine, and glutamic acid; B. subtilis exhibited the highest production of alanine, asparagine, glycine, leucine, proline, tryptophan, and lysine. B. velezensis increased the γ-aminobutyric acid (GABA) concentration to >200% of that in the control samples. B. sonorensis produced a somewhat similar amino acid profile with B. licheniformis. Comparative genomic analysis of the four Bacillus strains and the genetic profiles of the produced free amino acids revealed that genes involved in glutamate and arginine metabolism were not common to the four strains. The genes gadA/B (encoding a glutamate decarboxylase), rocE (amino acid permease), and puuD (γ-glutamyl-γ-aminobutyrate hydrolase) determined GABA production, and their presence was species-specific. Taken together, B. licheniformis and B. velezensis were respectively shown to have high potential to increase concentrations of glutamic acid and GABA, while B. subtilis has the ability to increase essential amino acid concentrations in fermented soybean foods.

Primary metabolic responses in the leaves and roots of bell pepper plants subjected to microelements-deficient conditions

  • Sung, Jwakyung;Lee, Yejin;Lee, Seulbi
    • Korean Journal of Agricultural Science
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    • v.48 no.1
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    • pp.179-189
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    • 2021
  • Plants need essential mineral elements to favorably develop and to complete their life cycle. Despite the irreplaceable roles of microelements, they are often ignored due to the relative importance of macroelements with their influence on crop growth and development. We focused on the changes in primary metabolites in the leaves and roots of bell pepper plants under 6 microelements-deficient conditions: Copper (Cu), Zinc (Zn), Iron (Fe), Manganese (Mn), Boron (B) and Molybdenum (Mo). Bell pepper plants were grown in hydroponic containers, and individual elements were adjusted to 1/10-strength of Hoagland nutrient solution. A remarkable perturbation in the abundance of the primary metabolites was observed for the Fe and B and the Mn and B deficiencies in the leaves and roots, respectively. The metabolites with up-accumulation in the Fe-deficient leaves were glucose, fructose, xylose, glutamine, asparagine and serine. In contrast, the Mn deficiency also resulted in a higher accumulation of glucose, fructose, xylose, galactose, serine, glycine, β-alanine, alanine and valine in the roots. The B deficiency noticeably accumulated alanine, valine and phenylalanine in the roots while it showed a substantial decrease in glucose, fructose and xylose. These results show that the primary metabolism could be seriously disturbed due to a microelement deficiency, and the alteration may be either the specific or adaptive responses of bell pepper plants.

Time-dependent changes of fruit metabolites studied by 1H NMR

  • Park, Sung Jean
    • Journal of the Korean Magnetic Resonance Society
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    • v.26 no.3
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    • pp.24-33
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    • 2022
  • The browning phenomenon of fruits can be easily observed when fruits or vegetables (apples, pears, bananas, potatoes, etc.) are cut with a knife and the part turns brown. When this browning occurs, changes in taste, color, and nutrients usually are introduced. The cause of this browning phenomenon has been well studied for a long time, but these studies have mainly focused on preventing deterioration of processed foods during food processing or storage. Resultantly, there are few studies on how much changes in nutrients (saccharides, amino acids, fats, water-soluble low molecular weight ammonium ions, etc.) are caused by browning. The purpose of this study is to determine the change in nutrients during browning using apple as a model fruit. We conducted a comparative study on how much the nutrient fluctuations differ depending on the presence or absence of pretreatment such as the application of heat. All analysis was conducted using 1H NMR. The ANOVA analysis showed that the concentrations of 4 amino acids (alanine, asparagine, isoleucine, and valine), 3 types of sugars (fructose, glucose, and xylose), 1 type of organic acid (lactate) and choline were significantly increased in samples showing browning. In addition, the groups before and after browning were clearly separated using multivariate statistical analysis methods (PCA, PLS-DA), which was greatly contributed by two sugar components (fructose and glucose) present in high concentrations in apples.

Aspartate-glutamate carrier 2 (citrin): a role in glucose and amino acid metabolism in the liver

  • Milan Holecek
    • BMB Reports
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    • v.56 no.7
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    • pp.385-391
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    • 2023
  • Aspartate-glutamate carrier 2 (AGC2, citrin) is a mitochondrial carrier expressed in the liver that transports aspartate from mitochondria into the cytosol in exchange for glutamate. The AGC2 is the main component of the malate-aspartate shuttle (MAS) that ensures indirect transport of NADH produced in the cytosol during glycolysis, lactate oxidation to pyruvate, and ethanol oxidation to acetaldehyde into mitochondria. Through MAS, AGC2 is necessary to maintain intracellular redox balance, mitochondrial respiration, and ATP synthesis. Through elevated cytosolic Ca2+ level, the AGC2 is stimulated by catecholamines and glucagon during starvation, exercise, and muscle wasting disorders. In these conditions, AGC2 increases aspartate input to the urea cycle, where aspartate is a source of one of two nitrogen atoms in the urea molecule (the other is ammonia), and a substrate for the synthesis of fumarate that is gradually converted to oxaloacetate, the starting substrate for gluconeogenesis. Furthermore, aspartate is a substrate for the synthesis of asparagine, nucleotides, and proteins. It is concluded that AGC2 plays a fundamental role in the compartmentalization of aspartate and glutamate metabolism and linkage of the reactions of MAS, glycolysis, gluconeogenesis, amino acid catabolism, urea cycle, protein synthesis, and cell proliferation. Targeting of AGC genes may represent a new therapeutic strategy to fight cancer.

Studies on Sclerotium rolfsii Sacc. isolated from Magnolia kobus DC. in Korea (목련(Magnolia kobus DC.)에서 분리한 흰비단병균(Sclerotium rolfsii Sacc.)에 관한 연구)

  • Kim Kichung
    • Korean journal of applied entomology
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    • v.13 no.3 s.20
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    • pp.105-133
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    • 1974
  • The present study is an attempt to solve the basic problems involved in the control of the Sclerotium disease. The biologic stranis of Sclerotium rolfsii Sacc., pathogen of Sclerotium disease of Magnolia kobus, were differentiated, and the effects of vitamins, various nitrogen and carbon sources on its mycelial growth and sclerotial production have been investigated. In addition the relationship between the cultural filtrate of Penicillium sp. and the growth of Sclerotium rolfsii, the tolerance of its mycelia or sclerotia to moist heat or drought and to Benlate (methyl-(butylcarbamoy 1)-2-benzimidazole carbamate), Tachigaren (3-hydroxy-5-methylisoxazole) and other chemicals were also clarified. The results are summarizee as follows: 1. There were two biologic strains, Type-l and Type-2 among isolates. They differed from each other in the mode of growth and colonial appearance on the media, aversion phenomenon and in their pathogenicity. These two types had similar pathogenicity to the Magnolia kobus and Robinia pseudoacasia, but behaved somewhat differently to the soybaen and cucumber, the Type-l being more virulent. 2. Except potassium nitrite, sodium nitrite and glycine, all of the 12 nitrogen sources tested were utilized for the mycelial growth and sclerotial production of this fungus when 10r/l of thiamine hydrochloride was added in the culture solution. Considering the forms of nitrogen, ammonium nitrogen was more available than nitrate nitrogen for the growth of mycelia, but nitrate nitrogen was better for sclerotia formation. Organic nitrogen showed different availabilities according to compounds used. While nitrite nitrogen was unavailable for both mycelial growth and sclerotial formation whether thiamine hydrochlioride was added or not. 3. Seven kinds of carbon sources examined were not effective in general, as long as thiamine hydrochloride was not added. When thiamine hydrochloride was added, glucose and saccharose exhibited mycelial growth, while rnaltose and soluble starch gave lesser, and xylose, lactose, and glycine showed no effect at all,. In the sclerotial production, all the tested carbon sources, except lactose, were effective, and glucose, maltose, saccharose, and soluble starch gave better results. 4. At the same level of nitrogen, the amount of mycelial growth increased as more carbon Sources were applied but decreased with the increase of nitrogen above 0.5g/1. The amount of sclerotial production decreased wi th the increase of carbon sources. 5. Sclerotium rolfsii was thiamine-defficient and required thiamine 20r/l for maximun growth of mycelia. At a higher concentration of more than 20r/l, however, mycelial growth decreased as the concentration increased, and was inhibited at l50r/l to such a degree of thiamine-free. 6. The effect of the nitrogen sources on the mycelial growth under the presence of thiamine were recognized in the decreasing order of $NH_4NO_3,\;(NH_4)_2SO_4,\;asparagine,\;KNO_3$, and their effects on the sclerotial production in the order of $KNO_3,\;NH_4NO_3,\;asparagine,\;(NH_4)_2SO_4$. The optimum concentration of thiamine was about 12r/l in $KNO_3$ and about 16r/l in asparagine for the growth of mycelia; about 8r/l in $KNO_3$ and $NH_4NO_3$, and 16r/l in asparagine for the production of sclerotia. 7. After the fungus started to grow, the pH value of cultural filtrate rapidly dropped to about 3.5. Hereafter, its rate slowed down as the growth amount increased and did not depreciated below pH2.2. 8. The role of thiamine in the growth of the organism was vital. If thiamine was not added, the combination of biotin, pyridoxine, and inositol did not show any effects on the growth of the organism at all. Equivalent or better mycelial growth was recognized in the combination of thiamine+pyridoxine, thiamine+inositol, thiamine+biotin+pyridoxine, and thiamine+biotin+pyridoxine+inositol, as compared with thiamine alone. In the combinations of thiamine+biotin and thiamine+biotin+inositol, mycelial growth was inhibited. Sclerotial production in dry weight increased more in these combinations than in the medium of thiamine alone. 9. The stimulating effects of the Penicillium cultural filtrate on the mycelial growth was noticed. It increased linearly with the increase of filtrate concentration up to 6-15 ml/50ml basal medium solution. 10. $NH_4NO_3$. as a nitrogen source for mycelial growth was more effective than asparasine regardless of the concentration of cultural filtrate. 11. In the series of fractionations of the cultural filtrate, mycelial growth occured in unvolatile, ether insoluble cation-adsorbed or anion-unadsorbed substance fractions among the fractions of volatile, unvolatile acids, ether soluble organic acids, ether insoluble, cation-adsorbed, cation-unadsorbed, anion-adsorbed and anion-unadsorbed. and anion-un-adsorbed substance tested. Sclerotia were produced only in cation-adsorbed fraction. 12. According to the above results, it was assumed that substances for the mycelial growth and sclerotial formation and inhibitor of sclerotial formation were include::! in cultural filtrate and they were quite different from each other. I was further assumed that the former two substances are un volatile, ether insotuble, and adsorbed to cation-exchange resin, but not adsorbed to anion, whereas the latter is unvolatile, ether insoluble, and not adsorbed to cation or anion-exchange resin. 13. Seven amino acids-aspartic acid, cystine, glysine, histidine, Iycine, tyrosine and dinitroaniline-were detected in the fractions adsorbed to cation-exchange resin by applying the paper chromatography improved with DNP-amino acids. 14. Mycelial growth or sclerotial production was not stimulated significantly by separate or combined application of glutamic acid, aspartic acid, cystine, histidine, and glysine. Tyrosine gave the stimulating effect when applied .alone and when combined with other amino acids in some cases. 15. The tolerance of sclerotia to moist heat varied according to their water content, that was, the dried sclerotia are more tolerant than wet ones. The sclerotia harvested directly from the media, both Type-1 and Type-2, lost viability within 5 minutes at $52^{\circ}C$. Sclerotia dried for 155 days at$26^{\circ}C$ had more tolerance: sclerotia of Type-l were killed in 15 mins. at $52^{\circ}C$ and in 5 mins. at $57^{\circ}C$, and sclerotia of Type-2 were killed in 10 mins. both at $52^{\circ}C$ or $57^{\circ}C$. 16. Cultural sclerotia of both strains maintained good germinability for 132 days at$26^{\circ}C$. Natural sclerotia of them stored for 283 days under air dry condition still had good germinability, even for 443 days: type-l and type-2 maintained $20\%$ and $26.9\%$ germinability, respectively. 17. The tolerance to low temperature increased in the order of mycelia, felts and sclerotia. Mycelia completely lost the ability to grow within 1 week at $7-8^{\circ}C$> below zero, while mycelial felts still maintained the viability after .3 weeks at $7-20^{\circ}C$ below zero, and sclerotia were even more tolerant. 18. Sclerotia of type-l and type-2 were killed when dipped into the $0.05\%$ solution of mercury chloride for 180 mins. and 240 mins. respectively: and in the $0.1\%$ solution, Type-l for 60 mins. and Type-2 for 30 mins. In the $0.125\%$ uspulun solution, Type-l sclerotia were killed in 180 mins., and those of Type-2 were killed for 90 mins. in the$0.125\%$solution. Dipping into the $5\%$ copper sulphate solution or $0.2\%$ solution of Ceresan lime or Mercron for 240 mins. failed to kill sclerotia of either Type-l or Type-2. 19. Inhibitory effect on mycelial growth of Benlate or Tachi-garen in the liquid culture increased as the concentration increased. 6 days after application, obvious inhibitory effects were found in all treatments except Benlate 0.5ppm; but after 12 days, distingushed diflerences were shown among the different concentrations. As compared with the control, mycelial growth was inhibited by $66\%$ at 0.5ppm and by $92\%$ at 2.0ppm of Benlate, and by$54\%$ at 1ppm and about $77\%$ at 1.5ppm or 2.0ppm of Tachigaren. The mycelial growth was inhibited completely at 500ppm of both fungicides, and the formation of sclerotia was checked at 1,000ppm of Benlate ant at 500ppm or 1,000ppm of Tachigaren. 20. Consumptions of glucose or ammonium nitrogen in the culture solution usually increased with the increment of mycelial growth, but when Benlate or Tachigaren were applied, consumptions of glucose or ammonium nitrogen were inhibited with the increment of concentration of the fungicides. At the low concentrations of Benlate (0.5ppm or 1ppm), however, ammonium nitrogen consumption was higher than that of the ontrol. 21. The amount of mycelia produced by consuming 1mg of glucose or ammonium nitrogen in the culture solution was lowered markedly by Benlate or Tachigaren. Such effects were the severest on the third day after their treatment in all concentrations, and then gradually recovered with the progress of time. 22. In the sand culture, mycelial growth was not inhibited. It was indirectly estimated by the amount of $CO_2$ evolved at any concentrations, except in the Tachigaren 100mg/g sand in which mycelial growth was inhibited significantly. Sclerotial production was completely depressed in the 10mg/g sand of Benlate or Tachigaren. 23. There was no visible inhibitory effect on the germination of sclerotia when the sclerotia were dipped in the solution 0.1, 1.0, 100, 1.000ppm of Benlate or Tachigaren for 10 minutes or even 20 minutes.

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Pepstatin- Insensitive Carboxyl Proteinase: A Biochemical Marker for Late Lysosomes in Amoeba proteus

  • Hae Kyung Kwon;HyeonJung Kim;Tae In Ahn
    • Animal cells and systems
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    • v.3 no.2
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    • pp.221-228
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    • 1999
  • In order to find a biochemical marker for late Iysosomes, we characterized two cDNAs which were cloned by using a monoclonal antibody (mAb) against Iysosomes in Amoeba proteus as a probe. The two cDNAs, a 1.3-kb cDNA in pBSK-Iys45 and a 1.6-kb cDNA in pBSK-Iys60, were found to encode proteins homologous to pepstatin-insensitive carboxyl proteinases (PICPs). E. coli transformed with pBSK-Iys45 produced two immunopositive polypeptides (45 and 43 kDa) and the cDNA in 1274 bases encoded a 44,733-Da protein (Lys45) of 420 amino acids containing one site for a core oligosaccharide. On the other hand, E. coli transformed with pBSK-Iys60 produced several polypeptides (64, 54, 45, 41, and 37 kDa) reacting with the mAb. The cDNA contained 1629 bases and encoded a 59,231-Da protein (Lys60) of 530 amino acids containing two sites for asparagine-linked core oligosaccharides. These two cDNAs showed identities of 60.3% in nucleotide sequences and 23.6% in amino acid sequences. Lys45 and Lys60 appeared to share XXEFQK as a common antigenic domain. The amino acid sequence of the Lys45 protein showed 17.4% identity and 40.9% similarity to that of PICP from Pseudomonas sp. 101. On the other hand, Lys60 showed a 24.3% identity and 51.9% similarity with human Iysosomal PICP in the amino acid sequence. A putative active center for serine protease, GTS*xxxxxFxG, was found to be conserved among PICP homologues. The two PICPs are the first reported enzymatic markers for late Iysosomes.

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