• Title/Summary/Keyword: Protein A

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Biochemical and Molecular Biological Studies on the DNA Replication of Bacteriophage T7 (Bacteriophage T7의 유전자 복제기작에 관한 생화학적, 분자생물학적 특성 연구)

  • KIM Young Tae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.28 no.2
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    • pp.209-218
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    • 1995
  • Bacteriophage T7 gene 2.5 protein, a single-stranded DNA binding protein, has been implicated in T7 DNA replication, recombination, and repair. Purified gene 2.5 protein has been shown to interact with the phage encoded gene 5 protein (DNA polymerase) and gene 4 proteins (helicase and primase) and stimulates their activities. Genetic analysis of T7 phage defective in gene 2.5 shows that the gene 2.5 protein is essential for T7 DNA replication and growth. T7 phage that contain null mutants of gene 2.5 were constructed by homologous recombination. These mutant phage $(T7\Delta2.5)$ cannot grow in Escherichia coli. After infection of E. coli with $T7\Delta2.5$, host DNA synthesis is shut off, and $T7\Delta2.5$ DNA synthesis is reduced to less than $1\%$ of wild-type phage DNA synthesis (Kim and Richardson, 1993, Proc. Natl. Aca. Sci. USA, 90, 10173-10177). A truncated gene 2.5 protein $(GP2.5-\Delta21C)$ deleted the 21 carboxyl terminal amino acids was constructed by in vitro mutagenesis. $GP2.5-\Delta21C$ cannot substitute for wild-type gene 2.5 protein in vivo; the phage are not viable and exhibit less than $1\%$ of the DNA synthesis observed in wild-type phage-infected cells. $GP2.5-\Delta21C$ has been purified to apparent homogeneity from cells overexpressing its cloned gene. Purified $GP2.5-\Delta21C$ does not physically into「act with T1 gene 4 protein as measured by affinity chromatography and immunoblot analysis. The mutant protein cannot stimulate T7 gene 4 protein activity on RNA-primed DNA synthesis and primer synthesis. These results suggest that C-terminal domain of gene 2.5 protein is essential for protein-protein interactions.

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Role of Protein Kinase C on Norepinephrine Induced Inhibition of Calcium Current in Rat Sympathetic Neurons (흰쥐 교감신경세포에서 Norepinephrine 에 의한 칼슘전류 억제에 미치는 Protein Kinase C 의 역할)

  • ;Keith S. Elmslie
    • Progress in Medical Physics
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    • v.11 no.1
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    • pp.29-38
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    • 2000
  • The signal transduction pathway for most neurotransmitter induced inhibition of $Ca^{2+}$ channels in sympathetic neurons involves a G-protein mediated, membrane-delimited mechanism without the participation of any known protein kinase. However, activation of protein kinase C (PKC) has been proposed as one of the intracellular mechanisms mediating some neurotransmitter induced $Ca^{2+}$ channel inhibition. In the present study, we investigated the effects of phorbol-12, 13-dibutyrate (PDBu) on $Ca^{2+}$ channel currents of acutely dispersed neurons from adult rat superior cervical ganglion (SCG) neurons using whole cell variant of the patch clamp technique. PDBu (500 nM), the activator of PKC, increased $Ca^{2+}$ channel currents and retarded the deactivation of tail currents. The effects of PDBu were voltage dependent and the maximal increase in the current amplitudes was observed between -10 to 10 mV (n=4). PDBu attenuated $Ca^{2+}$ current inhibition induced by norepinephrine (NE), which modulates $Ca^{2+}$ channels via a pertussis toxin (PTX)-sensitive pathway. Inhibition of PDBu by staurosporine (1 $\mu$M) blocked the effects of PDBu on current amplitudes and NE-induced G-protein mediated inhibition of $Ca^{2+}$ currents. Further experiment should be done to know if G-protein or $Ca^{2+}$ channel itself is the target of PKC phosphorvlation.phosphorvlation.

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Interaction of the Bacteriophage P2 Tin Protein and Bacteriophage T4 gp32 Protein Inhibites Growth of Bacteriophage T4

  • Jin, Hee-Kyung;Kim, Min-Jung;Park, Chan-Hee;Park, Jung-Chan;Myung, Hee-Joon
    • Journal of Microbiology and Biotechnology
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    • v.11 no.4
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    • pp.724-726
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    • 2001
  • The growth of baceriophage T4 is inhibited by the presence of the tin gene product o bacteriophage P2. The interaction between purified Tin and gp32 proteins was observed using coimmunoprecipitation experiments. The in vivo interaction was confirmed by yeast two-hybrid experiments. A deletion analysis showed that the Asp 163 region of gp32 to DNA substrates was not affected by the presence of Tin, Thus, it would appear that the inhibition of 4 growth by Tin was due to a protein-protein interaction rather than affecting the DNA-binding ability of gp32.

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Studies on the Developmental Processes of the Protein Body in the Ginseng(Panax ginseng C.A. Meyer) Endosperm Cell (인삼(Panax ginseng C.A. Meyer) 배유세포의 Protein Body 형성에 관한 연구)

  • Yu, Seong-Cheol;Jeong, Byung-Kap;Kim, Woo-Kap
    • Applied Microscopy
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    • v.18 no.2
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    • pp.141-152
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    • 1988
  • The developmental processes of the protein body are studied on endosperm cells of Panax ginseng during seed maturation periods. The spherosome, mitochondria, rough endoplasmic reticulum, and ribosome are observed and then are gradually increased in early endosperm cells. Protein body developed from vesicles produced by the rough endoplasmic reticulum and was formed at the enlarged ends of rough endoplasmic reticulum. Also, vacuole-like protein body was observed in associated with rough endoplasmic reticulum. Golgi complex is observed in associated with vacuole and its vesicles containing proteinaceous granules moved and accumulated to the vacuole. Proteinaceous granules are deposited in the spherical or oval shaped vacuole and gradually, vacuole is surrounded by the multi-membranous structure. Rough endoplasmic reticulum, ribosome, Golgi complex, and vacuole are observed in associated with protein body formation.

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A New Approach to Find Orthologous Proteins Using Sequence and Protein-Protein Interaction Similarity

  • Kim, Min-Kyung;Seol, Young-Joo;Park, Hyun-Seok;Jang, Seung-Hwan;Shin, Hang-Cheol;Cho, Kwang-Hwi
    • Genomics & Informatics
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    • v.7 no.3
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    • pp.141-147
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    • 2009
  • Developed proteome-scale ortholog and paralog prediction methods are mainly based on sequence similarity. However, it is known that even the closest BLAST hit often does not mean the closest neighbor. For this reason, we added conserved interaction information to find orthologs. We propose a genome-scale, automated ortholog prediction method, named OrthoInterBlast. The method is based on both sequence and interaction similarity. When we applied this method to fly and yeast, 17% of the ortholog candidates were different compared with the results of Inparanoid. By adding protein-protein interaction information, proteins that have low sequence similarity still can be selected as orthologs, which can not be easily detected by sequence homology alone.

Nanoscale Protein Chip based on Electrical Detection

  • Choi, Jeong-Woo
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.18-18
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    • 2005
  • Photoinduced electron transport process in nature such as photoelectric conversion and long-range electron transfer in photosynthetic organisms are known to occur not only very efficiently but also unidirectionally through the functional groups of biomolecules. The basic principles in the development of new functional devices can be inspired from the biological systems such as molecular recognition, electron transfer chain, or photosynthetic reaction center. By mimicking the organization of the biological system, molecular electronic devices can be realized $artificially^{1)}$. The nano-fabrication technology of biomolecules was applied to the development of nano-protein chip for simultaneously analyzing many kinds of proteins as a rapid tool for proteome research. The results showed that the self-assembled protein layer had an influence on the sensitivity of the fabricated bio-surface to the target molecules, which would give us a way to fabricate the nano-protein chip with high sensitivity. The results implicate that the biosurface fabrication using self-assembled protein molecules could be successfully applied to the construction of nanoscale bio-photodiode and nano-protein chip based on electrical detection.

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Partial Purification of Mussel Adhesive Protein for Mytilus Edulis and Preparation of Mussel Protein Hydrolysates

  • Sun, Nam-Kyu;Song, Kyung-Bin
    • Preventive Nutrition and Food Science
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    • v.5 no.3
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    • pp.148-152
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    • 2000
  • Mussel adhesive protein (MAP) was extracted from Korean Mytilus edulis and then partially purified using Sephacryl S-300 gel permeation chromatography and reversed-phase high performance liquid chromatography. As an indicator of adhesiveness, is 3,4-dihydroxyphenylalanine (DOPA) content was determined. Its DOPA/protein ratio of 0.19 was higher than those of other reports, indicating a good adhesive. The partially purified MAP was confirmed by acid-urea polyacrylamide gel electrophoresis using cetylpiridinium bromide as a cationic detergent. Sea mussel hydrolysates were prepared using three commercial proteases to provide value-added functional materials and their angiotensin converting enzyme (ACE) inhibitory activities were determined. Among hydrolysates of sea mussel, Protamex was the best and further purification would improved ACE inhibitory activity.

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Food Components of Coho Salmon and Rainbowtrout (은연어와 무지개 송어의 식품성분)

  • 김경삼;최영준
    • The Korean Journal of Food And Nutrition
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    • v.6 no.2
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    • pp.73-80
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    • 1993
  • In order to examine food components of coho salmon and rainbow-trout, We analyzed the composition of protein, amino acids and total lipids. The coho salmon muscle contained about 19.3% of protein with the composition of 29.9% in sarcoplasmic protein, 56.3oA in myofibrillar protein 12.5% alkali soluble protein and 2.6% in stroma. Those of rainbow-trout contained 34.1%, 56.4%, 8.3% and 2.9%, respectively. The sarcoplasmic and myofibrillar protein were composed of 13 subunits in coho salmon, and 16 and 15 subunits in rainbow-trout. Judging from the contents of essential amino acids, both muscle proteins were complete proteins. The most remarkable feature of free amino acids was that a large amount of dipeptide anserine was present with fairly lower levels of 1 methyl histidine, taurine, histidine, alanine and glycine in both muscle extracts. The total fatty acids of coho salmon was composed of 31.49% polyenes, 43.79% monoenes and 24.73% saturates. The composition of total fatty acid of coho salmon muscle was not different from that of rainbow-trout muscle.

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Backbone Dynamics and Model-Free Analysis of N-terminal Domain of Human Replication Protein A 70

  • Yoo, Sooji;Park, Chin-Ju
    • Journal of the Korean Magnetic Resonance Society
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    • v.22 no.1
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    • pp.18-25
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    • 2018
  • Replication protein A (RPA) is an essential single-stranded DNA binding protein in DNA processing. It is known that N terminal domain of RPA70 (RPA70N) recruits various protein partners including damage-response proteins such as p53, ATRIP, Rad9, and MRE11. Although the common binding residues of RPA70N were revealed, dynamic properties of the protein are not studied yet. In this study, we measured $^{15}N$ relaxation parameters ($T_1,\;T_2$ and heteronuclear NOE) of human RPA70N and analyzed them using model-free analysis. Our data showed that the two loops near the binding site experience fast time scale motion while the binding site does not. It suggests that the protein binding surface of RPA70N is mostly rigid for minimizing entropy cost of binding and the loops can experience conformational changes.

Protein Qualities of Seasoned Tofu Containing Freshwater Crab Meat (참게육 첨가 맛 두부의 단백질 품질)

  • Ryu, Hong-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.42 no.6
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    • pp.580-584
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    • 2009
  • The effect of freshwater crab meat additive on protein quality of tofu was studied. Tofu containing freshwater crab meat(TCM) prepared by the formulation for the best sensory qualities had about 20% more lipid and ash content than those of commercial tofu. TCM showed a higher content of lysine, methionine, cysteine and tyrosine compared with those in commercial tofu. TCM was comparable to commercial tofu(CT) regarding the in vitro protein digestibility(95%). A considerable difference in computed protein efficiency ratio(C-PER) resulted between CT(1.76-1.81) and TCM(2.61). Therefore, crab meat has potential as an ingredient for enriching essential amino acid and improving protein quality of CT.