• 제목/요약/키워드: hprK

검색결과 26건 처리시간 0.021초

Characterization of a Bifunctional HPr Kinase/Phosphorylase from Leuconostoc mesenteroides SY1

  • Park, Jae-Yong;Lee, Kang-Wook;Lee, Ae-Ran;Jeong, Woo-Ju;Chun, Ji-Yeon;Lee, Jong-Hoon;Kim, Jeong-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제18권4호
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    • pp.746-753
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    • 2008
  • The hprK gene encoding bifunctional HPrK/P (kinase/phosphorylase) was cloned from L. mesenteroides SY1, a strain isolated from kimchi. hprK was transcribed as a monocistronic gene. His-tagged HPrH16A and HPrK/P were produced in E. coli BL21 (DE3) using pET26b(+) and purified. HPrK/P phosphorylation assay with purified proteins showed that the kinase activity of HPrK/P increased at slightly acidic pHs. Divalent cations such as $Mg^{2+}$ and $Mn^{2+}$ and glycolytic intermediates such as fructose-1, 6-bisphosphate (FBP) and phosphoenolpyruvate (PEP) increased the kinase activity of HPrK/P, but inorganic phosphate strongly inhibited it. Kinetic studies for the kinase activity of HPrK/P showed that the apparent $K_m$ values were 0.18 and $14.57{\mu}M$ for ATP and HPr, respectively. The $K_m$ value for the phosphorylase activity of HPrK/P was $14.16{\mu}M$ for P-Ser-HPr (HPr phosphorylated at the serine residue).

우렁쉥이(Halocynthia roretzi) 유생의 뇌포 구조와 수압수용쳬의 형성 (Brain Vesicle Structure and Formation of the Hydrostatic Pressure Receptors in Larvae of the Ascidian (Halocynthia roretzi))

  • 김정은;서형주;김길중
    • 한국수산과학회지
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    • 제39권2호
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    • pp.94-99
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    • 2006
  • The tadpole larvae of most ascidians have two sensory pigment cells in their brain vesicle. The anterior otolith pigment cell is sensitive to gravity, whereas the posterior ocellus pigment cell responds to light. Besides these two sensory cells, the larvae also possess another type of sensory receptor cell: hydrostatic pressure receptor (Hpr) cells. The Hpr cells have been presumed to sense hydrostatic water pressure, although no functional analysis has been performed. In larvae of the ascidian Halocynthia reretzi, the development of the Hpr cells and their structure in the brain vesicle are poorly understood. To investigate the morphology and formation of the Hpr cells, we established a monoclonal antibody, Hpr-1, that specifically recognizes Hpr cells. The Hpr-1 antigens became detectable in the brain vesicle at the late tailbud stage. Each Hpr cell projected a small globular body, connected by a short stalk, into the lumen of the brain vesicle. The brain vesicle showed remarkable left-right asymmetry. Pigment cells were located on the right side in the lumen of the brain vesicle, whereas Hpr cells were present in the left side. After metamorphosis, the Hpr cells were observed near the rudimental siphons of the juvenile.

Regulation of Glycogen Concentration by the Histidine-Containing Phosphocarrier Protein HPr in Escherichia coli

  • Koo, Byung-Mo;Seok, Yeong-Jae
    • Journal of Microbiology
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    • 제39권1호
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    • pp.24-30
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    • 2001
  • In addition to effecting the catalysis of sugar uptake, the bacterial phosphoenolpyruvate::sugar phosphotransferase system regulates a variety of physiological processes. In a previous paper [Seok et al.,(1997) J. Biol. Chem. 272, 26511-26521], we reported the interaction with and allosteric regulation of Esiherichia coli glycogen phosphorylase activity by the histidine-containing phosphocarrier protein HPr in vitro. Here, we show that the specific interaction between HPr and glycogen phosphorylase occurs in vivo. To address the physiological role of the HPr-glycogen phosphorylase complex, intracellular glycogen levels were measured in E. coli strains transformed with various plasmids. While glycogen accumulated during the transition between exponential and stationary growth phases in wildtype cells, it did not accumulate in cells overproducing HPr or its inactive mutant regardless of the growth stage. From these results, we conclude that HPr mediates crosstalk between sugar uptake through the phosphoenolpyruvate:sugar phosphotransferase system and glycogen breakdown. The evolutionary significance of the HPr-glycogen phosphorylase complex is suggested.

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멍게의 수압수용체세포 형성에서 FGF와 MEK 신호의 역할 (Role of FGF and MEK Signaling in Formation of the Hydrostatic Pressure Receptor Cells during Ascidian Embryogenesis)

  • 서형주;김길중
    • 한국발생생물학회지:발생과생식
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    • 제13권4호
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    • pp.291-296
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    • 2009
  • 멍게 유생의 뇌포에는 2개의 감각색소세포인 평형기와 안점 이외에 또 다른 감각세포로 추정되는 수압수용체세포가 존재한다. 수압수용체세포 형성에 관해서는 현재까지 거의 알려진 것이 없다. 본 연구에서는 수압수용체세포 형성에서 FGF 신호전달 과정의 관련성을 조사했다. 수정란에 Hr-FGF9/16/20 antisense MO를 미세주입했을 때, 발생한 유생에서 수압수용체세포 특이적 Hpr-1 항원의 발현이 검출되지 않았다. 32세포기부터 FGF 수용체 억제제 SU5402 및 MEK 억제제 U0126을 처리한 배아도 수압수용체세포를 갖지 못한 유생으로 발생했다. 다음으로 수압수용체세포 형성에 FGF 신호전달 과정이 관련되는 시기를 자세히 조사했다. 수압수용체세포 형성에는 FGF 수용체 활성이 16세포기부터 64세포기까지 필요하다는 것이 시사되었다. U0126은 8세포기부터 후기 낭배기까지 Hpr-1 항원 발현을 억제했다. Hpr-1 항원 발현은 신경판기 직전부터 U0126의 영향을 받지 않았다. 따라서, 멍게에서 수압수용체세포 형성은 1차 신경유도기부터 후기 낭배기까지 FGF 신호전달 과정을 필요로 한다는 것이 밝혀졌다.

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Characterization of ptsHI Operon from Leuconostoc mesenteroides SY1, a Strain Isolated from Kimchi

  • Park Jae-Yong;Jeong Seon-Ju;Chun Ji-Yeon;Lee Jong-Hoon;Chung Dae-Kyun;Kim Jeong-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제16권6호
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    • pp.988-992
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    • 2006
  • The ptsHI operon from Leuconostoc mesenteroides ssp. mesenteroides SY1 (L. mesenteroides SY1), a strain isolated from kimchi, was cloned and characterized. The ptsH open reading frame (ORF) was 273 bp in size, which can encode a protein of 90 amino acid residues with a molecular weight of 9,212 Da. The pfsI ORF was 1,719 bp in size, which was capable of encoding a protein of 572 amino acids with a molecular mass of 62,549 Da. ptsH and pfsI genes were transcribed as a single transcript of 2.0 kb in size regardless of carbon sources, supporting the operon structure. Although the deduced amino acid sequences of the HPr and EI were highly homologous with those of other Gram-positive bacteria, an additional amino acid (glutamine at the $3^{rd}$ amino acid) was present in HPr from L. mesenteroides SY1. Phosphorylation sites of HPr included the histidine residue ($16^{th}$) and serine residue ($47^{th}$). Mutant HPrs, in which each phosphorylation site was mutated into alanine, were obtained, and phosphorylation with HPr and mutated HPrs showed that HPr was phosphorylated at the serine residue ($47^{th}$) by HPr kinaseiphosphorylase (HPr K/P).

분열효모에서 sphpr1 유전자의 결실이 생장 및 mRNA Export에 미치는 영향 (Effects of the Repression of sphpr1 Expression on Growth and mRNA Export in Fission Yeast)

  • 이현주;윤진호
    • 미생물학회지
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    • 제48권2호
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    • pp.171-174
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    • 2012
  • THOC1/Hpr1는 mRNA가 전사되는 동안 mRNP의 포장과 mRNA 방출에 관여하는 진화적으로 잘 보존된 THO 복합체의 한 소단위이다. 분열효모 Schizosaccharomyces pombe에서도 THOC1/Hpr1과 유사한 단백질을 암호화하는 유전자(sphpr1로 명명)를 찾아 그 특성을 조사하였다. 이배체 S. pombe 균주에 하나의 sphpr1 유전자만을 결실시킨 후 4분체 분석을 수행한 결과, 이 유전자는 생장에 필수적이었다. 티아민에 의해 발현이 조절되는 강력한 nmt1 프러모터를 이용하여 sphpr1를 과발현시키더라도 세포의 생장과 mRNA 방출에는 전혀 영향이 없었다. 하지만, sphpr1의 발현을 억제하면 생장이 억제되었으며 poly$(A)^+$ RNA가 핵 안에 축적되었다. 이와 같은 결과들은 sphpr1 유전자가 생장과 mRNA의 핵에서 세포질로의 방출에 관여하고 있음을 시사한다.

음식물쓰레기 이용 혐기 산발효에 의한 수소 및 유기산 생산: 축산폐수 첨가 효과 (Effect of Livestock Wastewater Addition on Hydrogen and Organic Acids Production Using Food Waste)

  • 장수진;김동훈;이모권;나정걸;김미선
    • 한국수소및신에너지학회논문집
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    • 제26권3호
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    • pp.199-205
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    • 2015
  • Organic wastes such as food waste (FW), livestock wastewater (LW), and sewage sludge (SWS) can produce hydrogen ($H_2$) by anaerobic acid fermentation. Expecially, FW which has high carbohydrate content produces $H_2$ and short chain fatty acids by indigenous $H_2$ producing microorganisms without adding inoculum, however $H_2$ production rate (HPR) and yield have to be improved to use a commercially available technology. In this study, LW was mixed to FW in different ratios (on chemical oxygen demand (COD) basis) as an auxiliary substrate. The mixture of FW and LW was pretreated at pH 2 using 6 N HCl for 12 h and then fermented at $37^{\circ}C$ for 28 h. HPR of FW, 254 mL $H_2/L/h$, was increased with the addition of LW, however, mixing ratio of LW to FW was reversely related to HPR, exhibiting HPR of 737, 733, 599, and 389 mL $H_2/L/h$ at the ratio of FW:LW=10:1, 10:2, 10:3, and 10:4 on COD basis, respectively. Maximum HPR and $H_2$ production yield of 737 $H_2/L/h$ and 1.74 mol $H_2/mol$ hexoseadded were obtained respectively at the ratio of FW:LW=10:1. Butyrate was the main organic acid produced and propionate was not detected throughout the experiment.

Interaction for Phosphotransfer between N-Terminal Domain of Enzyme I and HPr of E. coli Phosphoenolpyruvate:Sugar Phosphotransferase System.

  • Seok, Yeong-Jae
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 1998년도 학술발표회
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    • pp.12-12
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    • 1998
  • The interaction between the N-terminal domain of enzyme I (EIN) and the histidine-containing phosphocarrier protein HPr of the Escherichia coli phosphoenolpyruvate:sugar phosphotransferase system has been investigated by Isothermal Titration Calorimetry and heteronuclear magnetic resonance spectroscopy.(omitted)

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Growth, Blood Metabolites, and Health of Holstein Calves Fed Milk Replacer Containing Different Amounts of Energy and Protein

  • Lee, H.J.;Khan, M.A.;Lee, W.S.;Kim, H.S.;Ki, K.S.;Jang, S.J.;Hur, T.Y.;Khan, M.S.;Choi, Y.J.
    • Asian-Australasian Journal of Animal Sciences
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    • 제21권2호
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    • pp.198-203
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    • 2008
  • This study was conducted to compare the effects of feeding high protein and low energy milk replacer (HPR; CP 25%, ME 3.6 Mcal/kg DM) with low protein and high energy milk replacer (HPR; CP 21%, ME 4.2 Mcal/kg DM) on feed consumption, body weight (BW) gain, health and selected blood metabolites in Holstein calves during the pre-weaning period. At each feeding, each milk replacer (MR) was prepared by mixing 0.125 kg of dry MR in 1L of warm ($60^{\circ}C$) water. The calves were fed either HPR (n = 10) or HER (n = 10) using mobile plastic bottles fitted with soft rubber nipples. All calves received 1.8L diluted MR at each feeding 3 times daily during the first 4 weeks of age; feeding frequency was reduced to 2 times daily for the next 2 weeks of age and then to once daily during the last week of the experiment. Jugular blood was sampled in calves at day 7, 14, 21, 35 and 49 of age to enumerate selected metabolites. Daily MR, starter and hay intake during the pre-weaning period were similar in calves fed HPR and HER. Consumption of starter, MGH and total DM steadily increased with the age of calves. Final BW, daily BW gain and feed efficiency of calves were not affected by treatments. Serum glucose, cholesterol, creatinine were decreased (p<0.05) and blood urea N was increased (p<0.05) in calves fed HER or HPR as they grew older. Serum glucose, total protein and albumin concentrations in calves were not affected by treatments. Serum GPT and GOT concentrations were higher (p<0.05) in calves on HPR than on HER. Scouring score, days scoured, respiratory score, rectal temperature and general appearance were similar in calves fed HPR and HER. Poor general appearance (dullness and droopy ears) of calves fed either HPR or HER reflected nutritional insufficiency and stress. In conclusion, energy and protein concentrations in MR did not affect feed intake and BW gain in Holstein calves during the pre-weaning period. Poor general appearance and lower BW gain of calves compared to those reported in the literature for milk fed calves prompt a demand for further research to improve the daily nutrient supply to MR-fed calves.

주의력결핍/과잉운동장애(ADHD) 아동의 진단도구로서 부모용 행동 평가지의 타당도 연구 - 한국아동인성검사와 아동 ${\cdot}$ 청소년 행동평가척도를 중심으로 - (A VALIDITY STUDY OF PARENT BEHAVIORAL RATING SCALES AS DIAGNOSTIC TOOLS OF ATTENTION DEFICIT/HYPERACTIVITY DISORDER)

  • 김지혜;소유경;정유숙;이임순;홍성도
    • Journal of the Korean Academy of Child and Adolescent Psychiatry
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    • 제11권2호
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    • pp.282-289
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    • 2000
  • 본 연구에서는 현재 주의력-결핍 및 과잉운동장애(Attention Deficit/Hyperactivity Disorder, 이하 ADHD)의 주요한 평가척도 중의 하나로서 이용되고 있는 한국아동인성검사(Korean Personality Inventory for Children, K-PIC)와 아동 ${\cdot}$ 청소년 행동평가척도(Korean Child Behavior Checklist KCBCL)의 임상적 타당도에 대해 살펴보고자 하였다. 피검자들은 ADHD의 하위 유형 중 주의력-결핍 우세형(ADHD-I)으로 진단된 환아 11명과 과잉 행동-충동 우세형 및 복합형(ADHD-H)로 진단된 환아 23명, 우울, 불안 등 정서적 문제로 진단(신경증 집단;N) 받은 정신과 외래 환아 16명, 정상아동 15명 등 총 65명이었다. 먼저 각 집단별로 분산분석을 실시하였고, 각각 행동 평정 척도의 변별력을 측정하기 위하여 판별분석을 실시하였다. 이 집단들의 K-PIC와 K-CBCL 결과들에 대한 전체(omnibus) F-검증 결과, K-PIC의 과잉행동(HPR) 척도와 K-CBCL의 주의집중문제(Attention Problems, 이하 AP) 척도들은 분산분석결과 유의한 차이를 보였다. 하지만 사후검증 결과, ADHD-I 집단과 ADHD-H 집단 그리고 신경증 집단 사이에 유의한 차이를 보이지 않는 반면, 그 세 집단은 모두 정상집단과 .05 유의도 수준 하에서 유의한 차이를 나타내었다. 이러한 결과는 K-PIC의 과잉행동(HPR) 척도와 K-CBCL의 주의집중문제(Attention Problems) 척도의 경우, 기본적으로 임상군과 정상군을 구별하는데는 유용하지만 임상군내에서의 세부 진단을 위해 사용하는 데는 한계가 있음을 시사해준다. 또한 본 연구 결과 상대적으로 HPR 척도는 ADHD-H type을 감별하는 데 더 유용한 반면, K-CBCL의 AP 척도는 ADHD-I type을 감별하는 데 더 큰 강점을 갖는 것으로 보이나, HPR척도와 AP 척도 중 어느 하나만을 사용하는 것보다는 병행하여 사용하는 것이 더 바람직한 것으로 보인다. 특히 cut-off score 면에서 두 척도 중 어느 하나에서 T 점수 65점 이상 상승할 때 ADHD로 진단내리는 경우, 가장 효과적으로 정분류율(true positive rate;sensitivity)과 정배제율(true negative rate;specificity)을 조화시킬 수 있는 결정규칙이 되는 것으로 나타났다.

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