• Title/Summary/Keyword: iron binding protein

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Production of Iron-Binding Peptides from Colostral Whey by Enzymatic Hydrolysis

  • Kim, Sang-Bum;Ku, Min-Jung;Cho, Won-Mo;Ki, Kwang-Seok;Kim, Hyeon-Shup;Nam, Myoung-Soo
    • Food Science of Animal Resources
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    • v.30 no.6
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    • pp.923-929
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    • 2010
  • Colostral whey prepared from colostrum (pooled from first six post-partum milkings) was heated for 10 min at $100^{\circ}C$ Heated colostral whey was incubated with 1% enzymes (protein equivalent basis) for 15, 30, 60, 90, and 120 min at $50^{\circ}C$. Papain, pepsin, trypsin, and alcalase produced different degrees of hydrolysis (DH), 10.66%, 12.42%, 10.83%, and 25.31%, respectively, at an incubation time of 120 min. The SDS-PAGE reveals that significant amounts of bovine serum albumin (BSA), ${\beta}$-lactoglobulin (${\beta}$-LG), and ${\alpha}$-lactalbumin (${\alpha}$-LA) survived papain digestion. In contrast, pepsin completely removed BSA but not ${\beta}$-LG present in heated colostral whey. Alcalase completely eliminated BSA, ${\beta}$-LG, and ${\alpha}$-LA. This differential hydrolysis was confirmed by reversed-phase HPLC analysis. Using ion-exchange chromatography, fraction-1 (F-1) was obtained from alcalase hydrolysate at a NaCl gradient concentration of 0.25 M. Reversed-phase HPLC chromatograms of alcalase F-1 showed numerous small peaks, which probably indicate that a variety of new peptides were produced. Iron content of alcalase F-1 was 28.94 ppm, which was the highest among all enzyme fractions, whereas iron content of colostral whey was 36.56 ppm. Main amino acids contained in alcalase F-1 were Thr (15.45%), Glu (14.12%), and Ser (10.39%). Therefore, alcalase can be used to generate good iron-binding peptides in heated colostral whey, and the resulting iron-binding peptides could be suitable as a value-added food ingredient for food supplements.

Backbone NMR chemical shift assignment for the substrate binding domain of Escherichia coli HscA

  • Jin Hae Kim
    • Journal of the Korean Magnetic Resonance Society
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    • v.28 no.2
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    • pp.6-9
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    • 2024
  • HscA is a Hsp70-type chaperone protein that plays an essential role to mediate the iron-sulfur (Fe-S) cluster biogenesis mechanism in Escherichia coli. Like other Hsp70 chaperones, HscA is composed of two domains: the nucleotide binding domain (NBD), which can hydrolyze ATP and use its chemical energy to facilitate the Fe-S cluster transfer process, and the substrate binding domain (SBD), which directly interacts with the substrate, IscU, the scaffold protein of an Fe-S cluster. In the present work, we prepared the isolated SBD construct of HscA (HscA(SBD)) and conducted the solution-state nuclear magnetic resonance (NMR) experiments to have its backbone chemical shift assignment information. Due to low spectral quality of HscA(SBD), we obtained all the NMR data from the sample containing the peptide LPPVKIHC, the HscA-interaction motif of IscU, from which the chemical shift assignment could be done successfully. We expect that this information provides an important basis to execute detailed structural characterization of HscA and appreciate its interaction with IscU.

Regulation of Ferritin Synthesis by Iron-responsive Element in 5'-Untranslated Region (5'-Untranslated Region에 존재하는 Iron Responsive Element에 의한 Ferritin 합성조절)

  • Chung, In-Sik;Lee, Jung-Lim;Kim, Hae-Yeong
    • Applied Biological Chemistry
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    • v.41 no.3
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    • pp.224-227
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    • 1998
  • The expression of ferritin involved in iron metabolism is regulated at the translational level by the interaction of iron regulatory protein with iron-responsive element(IRE) in the 5'-untranslated region of ferritin transcript. To identify the role of structural element utilized for translational regulation of ferritin, we studied the effects of mutations in the ferritin IRE by measuring IRP binding activity and translational activity. Our data suggest that the cytosine at bulged position of IRE within ferritin is important for the formation of RNA secondary structure involved in translational regulation.

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Staphylococcus aureus Siderophore-Mediated Iron-Acquisition System Plays a Dominant and Essential Role in the Utilization of Transferrin-Bound Iron

  • Park Ra Young;Sun Hui Yu;Choi Mi Hwa;Bai Young Hoon;Shin Sung Heui
    • Journal of Microbiology
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    • v.43 no.2
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    • pp.183-190
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    • 2005
  • Staphylococcus aureus is known to be capable of utilizing transferrin-bound iron, via both siderophore­and transferrin-binding protein (named IsdA)-mediated iron-acquisition systems. This study was designed in order to determine which iron-acquisition system plays the essential or dominant role with respect to the acquisition of iron from human transfenin, in the growth of S. aureus. Holotransferrin (HT) and partially iron-saturated transferrin (PT), but not apotransferrin (AT), were found to stimulate the growth of S. aureus. S. aureus consumed most of the transferrin-bound iron during the exponential growth phase. Extracellular proteases were not, however, involved in the liberation of iron from transferrin. Transferrin-binding to the washed whole cells via IsdA was not observed during the culture. The expression of IsdA was observed only in the deferrated media with AT, but not in the media supplemented with PT or HT. In contrast, siderophores were definitely produced in the deferrated media with PT and HT, as well as in the media supplemented with AT. The siderophores proved to have the ability to remove iron directly from transferrin, but the washed whole cells expressing IsdA did not. In the bioassay, the growth of S. aureus on transferrin-bound iron was stimulated by the siderophores alone. These results demonstrate that the siderophore-mediated iron-acquisition system plays a dominant and essential role in the uptake of iron from transferrin, whereas the IsdA-mediated iron-acquisition system may play only an ancillary role in the uptake of iron from transferrin.

Iron Status in Female College Students in the Gyeongnam Area (경남 일부지역 여대생의 철 영양상태에 관한 연구)

  • Park, Mi-Young;Kim, Sung-Hee
    • Journal of Nutrition and Health
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    • v.44 no.3
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    • pp.222-230
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    • 2011
  • This study was undertaken to investigate iron status and related factors in female college students residing in Gyeongnam. The subjects were divided into normal (40.8%) and iron deficiency (ID) groups (59.2%) by iron status. Mean height, weight, lean body mass, percent body fat, body mass index, and wrist to hip ratio were not significantly different between the groups, but basic metabolic rate was significantly higher in the normal group than that in the ID group. The levels of hemoglobin, hematocrit, serum ferritin, transferrin saturation, mean corpuscular volume, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration were significantly higher in the normal group than those in the ID group. However, total iron binding capacity was significantly lower in the normal group than that in the ID group. Daily intake of protein, heme-Fe, niacin, and vitamin C were significantly higher in the normal group than those in the ID group. The mean intake of protein, Fe, niacin, vitamin $B_{12}$, and vitamin C based on the Korean recommended intake (RI) were significantly higher in the normal group than those in the ID group. The mean intakes of Ca, vitamin $B_{12}$, and folate in both groups were < 75% of the Korean RI. In conclusion, increasing dietary heme-Fe and vitamin C may be helpful for preventing ID anemia in female college students.

Physical Performance in Young Women with Depleted Iron Stoles (젊은 여성의 철분저장 고갈상태와 운동수행능력)

  • 정영진;정명일;정종귀
    • Journal of Nutrition and Health
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    • v.35 no.2
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    • pp.223-228
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    • 2002
  • The purpose of this study was to investigate the relationship between iron deficiency without anemia and physical performance in healthy women aged 20-21 yrs. Ten subjects with normal iron stores (serum ferritin $\geq$ 12$\mu\textrm{g}$/L: iron-sufficient group) and 11 subjects with iron depletion without anemia (serum ferritin < 12 $\mu\textrm{g}$/L and serum hemoglobin > 120 g/L: iron-depleted group) were chosen from a group of 50 women and were given physical-performance tests, including determinations of maximum oxygen consumption (VO$_2$ max) and ventilatory threshold. Iron status assessment included determination of hemoglobin, hematocrit, seam ferritin, total iron-binding rapacity, serum iron and transferrin saturation values. Dietary iron intake was assessed based on seven-day food intake records written by the subjects. Physical activity level was estimated by frequency questionnaires and two-week physical activity records were compiled daily by the subjects. Blood ferritin concentration was significantly lower in the iron-depleted group than in the iron-sufficient group (p < 0.05). However, other variables showing iron status was not different between the groups. There were no significant differences in body size, body composition and physical activity levels between the groups. Daily dietary iron, total protein and animal protein intakes of the iron-sufficient group were significantly higher than those of the iron depleted group. However, no differences were found in the amount of dietary vitamin C and fiber between the groups. The values for VO$_2$max and VO$_2$max corrected with weight or fat-free mass were not different between the groups. However, the ventilatory threshold was significantly higher in the iron sufficient group than in the iron-depleted group. The lower ventilatory threshold in the iron-depleted group suggests that iron depletion without anemia could diminish aerobic physical performance in young women. In addition, a significant correlation of physical performance to serum fferritin level was shown only in the iron depleted group.

Regulation of Macrophage Ceruloplasmin Gene Expression: One Paradigm of 3'-UTR-mediated Translational Control

  • Mazumder, Barsanjit;Sampath, Prabha;Fox, Paul L.
    • Molecules and Cells
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    • v.20 no.2
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    • pp.167-172
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    • 2005
  • Ceruloplasmin (Cp) is a copper protein with important functions in iron homeostasis and in inflammation. Cp mRNA expression is induced by interferon (IFN)-${\gamma}$ in U937 monocytic cells, but synthesis of Cp protein is halted after about 12 h by transcript-specific translational silencing. The silencing mechanism requires binding of a 4-component cytosolic inhibitor complex, IFN-gamma-activated inhibitor of translation (GAIT), to a defined structural element (GAIT element) in the Cp 3'-UTR. Translational silencing of Cp mRNA requires the essential proteins of mRNA circularization, suggesting that the translational inhibition requires end-to-end mRNA closure. These studies describe a new mechanism of translational control, and may shed light on the role that macrophage-derived Cp plays at the intersection of iron homeostasis and inflammation.

Action Mechanism of LB10522, a New Catechol-Substituted Cephalosporin (카테콜 치환체를 가진 세파로스포린계 항생제 LB10522의 작용기전)

  • Kim, Mu-Yong;Oh, Jeong-In;Paek, Kyoung-Sook;Kim, In-Chull;Kwak, Jin-Hwan
    • YAKHAK HOEJI
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    • v.40 no.1
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    • pp.102-111
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    • 1996
  • LB10522 is a new parenteral broad spectrum cephalosporin with a catechol moiety at C-7 position of beta-lactam ring. This compound can utilize tonB-dependent iron transp ort system in addition to porin proteins to enter bacterial periplasmic space and access to penicillin-binding proteins (PBPs) which are the lethal targets of ${\beta}$-lactam antibiotics. The chelating activity of LB10522 to metal iron was measured by spectrophotometrically scanning the absorbance from 200 to 900nm. When $FeCl_3$ was added, optical density was increased between 450 and 800nm. LB10522 was more active against gram-negative strains in iron-depleted media than in iron-replete media. This is due to the increased expression of iron transport channels in iron-depleted condition. LB10522 showed a similar activity against E. coli DC2 (permeability mutant) and E. coli DCO (wild type strain) in both iron-depleted and iron-replete media, indicating a minimal permeaility barrier for LB10522 uptake. LB10522 had high affinities to PBP 3 and PBP 1A, 1B of E. coli. By blocking these proteins, LB10522 caused inhibition of cell division and the eventual death of cells. This result was correlated well with the morphological changes in E. coli exposed to LB10522. Although the in vitro MIC of LB10522 against P. aeruginosa 1912E mutant (tonB) was 8-times higher than that of the P. aeruginosa 1912E parent strain, LB10522 showed a similar in vivo protection efficacy against both strains in the mouse systemic infection model. This result suggested that tonB mutant, which requires a high level of iron for normal growth, might have a difficulty in surviving in their host with an iron-limited environment.

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A Study on Nutritional Status of Iron in the Rural Elderly (일부 지역 농촌 노인의 철분 영양상태에 관한 연구)

  • 신소영;최미경;이윤식;승정자
    • Journal of the East Asian Society of Dietary Life
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    • v.6 no.2
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    • pp.135-142
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    • 1996
  • The purpose of this study was to examine nutritional status of iron in 27 rural elderly subjects. The daily intake, urinary excretion and serum level of iron were determined by 24-hr food collection method, and 24-hr urine and serum collection for 3 days. The results were summarized as follows. 1. The mean age, height, weight, and blood pressure were 67.7 years, 163.2cm, 55.9kg, and 121.1/80. 0mmHg in men and 69.9 years, 146.5cm, 47.6kg, and 108.9/71.1mmHg in women, respectively 2. The daily intake of energy, protein, vitamin A, vitamin B$_1$, vitamin B$_2$, niacin, vitamin C, and iron were lower than Korean recommended dietary allowances. 3. The daily iron intake was 7.82mg. The proportion of heme iron and nonheme iron to the total iron intake were 8.05% and 91.94%, respectively Major iron sources of the rural elderly were cooked rice, Ko Chu Jang, beef blood, anchovy, soybean paste, and leafy radish. 4. The hemoglobin, hematocrit, serum iron, and total iron binding capacity were 14.3g/㎗, 42.9%, 79.4 $\mu\textrm{g}$/㎗, and 268.4 $\mu\textrm{g}$/㎗ in men and 12.9g/㎗, 38.8%,92.4 $\mu\textrm{g}$/㎗, and 277.9$\mu\textrm{g}$/㎗ in women, respectively. 5. There was a significant positive correlation between dietary intake and urinary excretion of iron.

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A Survey of Nutrition and Blood Contents of Rural Women in Korea (일부 농촌지역 부인의 영양실태 및 혈액성상에 관한 연구)

  • Lim, Hyeon-Sook;Hwang, Geum-Hee
    • Journal of Nutrition and Health
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    • v.15 no.3
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    • pp.171-180
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    • 1982
  • The nutritional survey and blood sampling was conducted from May 23 to 27, 1971, in order to investigate the state of the nutritional intake in relation to blood contents of rural women. This survey covered a total of 47 women residing in Youngam-gun, Cholla-namdo, Korea. The daily diet of these subjects was observed by 24-hour recall method and weighing method for 3 days and hemoglobin (Hb), hematocrit(Hct), mean corpuscular hemoglobin concentration (MCHC), serum protein, serum albumin, A/G ratio, serum iron, total iron binding capacity (TIBC), transferrin saturation (TS), serum cholesterol and blood specific gravity (SG) were measured. The results were summarized as follows ; The rate of intake of vegetable foods was high, especially cereals, and that of animal foods was low, especially eggs and milk and milk products. The daily intake of niacin was sufficient, and the intakes of energy and iron were also tend to be adequate. On the contrary, the intakes of vitamin A, $B_2$ and C, calcium and protein were insufficient. The mean red blood cell number was $410.8{\pm}26.4{\times}10^{4}/mm^{3},$ the mean values of Hb, Hct and MCHC were $12.5{\pm}0.9gm%$, $37.5{\pm}2.4%$ and $33.2{\pm}0.5%$ thus the anemic prevalences were 21.3%, 12.8% and 93.6% comparing with the anemic criteria of WHO. The levels of serum protein and serum albumin were $7.1{\pm}0.4gm\;%$ and $4.3{\pm}0.3gm\;%$so the A/G ratio was indicated $1.6{\pm}0.2.$. The levels of serum iron and TIBC were $91.9{\pm}27.5{\mu}g/100ml and $297.5{\pm}31.4{\mu}g/100ml,$ so TS was calculated $31.2{\pm}9.7\;%$. The mean value of SG was $1.044{\pm}0.002,$ and the level of serum cholesterol was $164.3{\pm}25.4mg/dl$. In the relation between each of bloood content, Hb showed positive correlation not only with Hct but also protein at the level of significance of 1 per cent and 5 per cent, and protein also showed a positive correlation with serum cholesterol at the 1 per cent level of significance. In the relation between the nutrient intake and blood contents, vitamin C intake showed a positive correlation with Hb, serum protein and serum iron at the level of significance of 1 per cent, iron intake also showed a positive correlation with serum iron and serum cholesterol at the same significant level, and protein intake showed a positive correlation with serum iron at the at the level of significance of 5 per cent.

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