• Title/Summary/Keyword: Protein A

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Presence of an Albumin-Like Protein on the Plasma Membrane of Adipocytes Isolated from Korean Cow

  • Choi, Chang-Bon;Rhee, Seung-Keun
    • BMB Reports
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    • v.31 no.6
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    • pp.542-547
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    • 1998
  • The main function of adipocytes in various species is to store nutrient energy in the form of triglycerides, and this function may he closely related with hormonal signaling through the plasma membrane of adipocytes. Using SDS-PAGE, two-dimensional gel electrophoresis, and a membrane biotinylation technique, we have identified a 55KDa protein (55K protein) from the plasma membrane fraction of adipocytes, with an isoelectric point (pI) of 8.1-8.3. However, this 55K protein was not observed with a two-dimensional gel electrophoresis carried out on plasma membrane fractions prepared from the liver, heart, and kidney tissues. An analysis of the 12 amino acids sequence at the N-terminal of the 55K protein showed that it has a similar sequence to that of bovine serum albumin.

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Expression of a Recombinant Cry1Ac Crystal Protein Fused with a Green Fluorescent Protein in Bacillus thuringiensis subsp. kurstaki $Cry^-B$

  • Roh Jong Yul;Lee In Hee;Li Ming Shun;Chang Jin Hee;Choi Jae Young;Boo Kyung Saeng;Je Yeon Ho
    • Journal of Microbiology
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    • v.42 no.4
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    • pp.340-345
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    • 2004
  • To investigate the co-expression and crystallization of a fusion gene between the Bacillus thuringiensis crystal protein and a foreign protein in B. thuringiensis, the expression of the Cry1Ac fused with green fluorescent protein (GFP) genes in a B. thuringiensis $Cry^-B$ strain was examined. The cry1Ac gene was cloned in the B. thuringiensis-E. coli shuttle vector, pHT3101, under the control of the native cry1Ac gene promoter, while the GFP gene was inserted into the XhoI site upstream of the proteolytic cleavage site, in the middle region of the crylAc gene (pProAc-GFP). The B. thuringiensis $Cry^-B$ strain carrying pProAc-GFP (ProAc-GFP/CB) did not produce any inclusion bodies. However, the transformed strain expressed fusion protein forms although the expression level was relatively low. Furthermore, an immu­noblot analysis using GFP and Cry1Ac antibodies showed that the fusion protein was not a single spe­cies, but rather multiple forms. In addition, the N-terminal fragment of Cry1Ac and a non-fused GFP were also found in the B. thuringiensis $Cry^-B$ strain after autolysis. The sporulated cells before autolysis and the spore-crystal mixture after autolysis of ProAc-GFP/CB exhibited insecticidal activities against Plutella xylostella larvae. Accordingly, the current results suggest that a fusion crystal protein produced by the transfomant, ProAc-GFP/CB, can be functionally expressed but easily degraded in B. thuring­iensis.

Expression of PACT and EIF2C2, Implicated in RNAi and MicroRNA Pathways, in Various Human Cell Lines

  • Lee, Yong-Sun;Jeon, Yesu;Park, Jong-Hoon;Hwang, Deog-Su;Dutta, Anindya
    • Animal cells and systems
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    • v.8 no.3
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    • pp.213-220
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    • 2004
  • MicroRNA and siRNA (small interfering RNA), representative members of small RNA, exert their effects on target gene expression through association with protein complexes called miRNP (microRNA associated ribonucleoproteins) and RISC (RNA induced silencing complex), respectively. Although the protein complexes are yet to be fully characterized, human EIF2C2 protein has been identified as a component of both miRNP and RISC. In this report, we raised antiserum against EIF2C2 in order to begin understanding the protein complexes. An immunoblot result indicates that EIF2C2 protein is ubiquitously expressed in a variety of cell lines from human and mouse. EIF2C2 protein exists in both cellular compartments, as indicated by an immunoblot assay with a nuclear extract and a cytosolic fraction (S100 fraction) from HeLa S3 lysate. Depletion of EIF2C1 or EIF2C2 protein resulted in a decrease of microRNA, suggesting a possible role of these proteins in microRNA stability or biogenesis. We also prepared antiserum against dsRNA binding protein PACT, whose homologs in C. elegans and Drosophila are known to have a role in the RNAi (RNA interference) pathway. The expression of PACT protein was also observed in a wide range of cell lines.

Glyphosate Toxicity: III. Detection of QB Protein in Thylakoid Membrane of Tomato Apical Meristem Using an Antibody Raised from Hybrid Protein of psb A and lac Z Gene (Glyphosate 독성: III. psb A와 lac Z 유전자의 Hybrid 단백질로부터 만들어진 항체를 이용한 토마토 정단분열조직의 Thylakoid막 내 QB 단백질의 검정)

  • Kim, Tae-Wan;Amrhein, Nikolaus
    • Korean Journal of Weed Science
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    • v.15 no.3
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    • pp.206-213
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    • 1995
  • Glyphosate(N-[phosphonomethyl]glycine) applied to the assimilate-exporting leaves(i.e. third old leaf) of tomato(Lycopersicon esculentum Mil var. Moneymaker). Herbicide binding protein, QB protein(D1), has been immunoblotted using the antibodies raised against the hybrid-protein expressed by a part of spinach psb A gene cloned in frame with the 3'end of lac Z gene to allow expression of the ${\beta}$-galactosidase(EC 3.21.23) in Escherichia coli. Glyphosate has an effect on a turnover of D1 within photosystem II of thylakoid membrane. The dysfunction of D1 protein within light harvesting complex(LHC-II) seems to be a pleiotropic effect of glyphosate.

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Protein-protein Interaction Analysis of Bradykinin Receptor B2 with Bradykinin and Kallidin

  • Nagarajan, Santhosh Kumar;Madhavan, Thirumurthy
    • Journal of Integrative Natural Science
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    • v.10 no.2
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    • pp.74-77
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    • 2017
  • Bradykinin receptor B2 (B2R) is a GPCR protein which binds with the inflammatory mediator hormone bradkynin. Kallidin, a decapeptide, also signals through this receptor. B2R is crucial in the cross-talk between renin-angiotensin system (RAS) and the kinin-kallikrein system (KKS) and in many processes including vasodilation, edema, smooth muscle spasm and pain fiber stimulation. Thus the structural study of the receptor becomes important. We have predicted the peptide structures of Bradykinin and Kallidin from their amino acid sequences and the structures were docked with the receptor structure. The results obtained from protein-protein docking could be helpful in studying the B2R structural features and in the pathophysiology in various diseases related to it.

Protein Delivery System based on Various Polysaccharides

  • Jung, Young-Seok;Na, Kun
    • Journal of Pharmaceutical Investigation
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    • v.41 no.4
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    • pp.197-204
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    • 2011
  • Delivery of therapeutic protein drugs is a hot issue in the clinical application, because protein drugs have low side effects and highly therapeutic effects compared with chemical drugs. Despite their prominent advantages, protein drugs have high risk for human therapy such as their easy degradation by proteolytic enzymes, renal filtration and immune response. Over the past few decades, a large number of polysaccharides as vehicles for the protein delivery system have been developed to overcome the problems. This review presents the studies on protein delivery based on polysaccharides used as stabilizer and vehicles comprising nano- or microspheres to overcome inherent limitations of therapeutic proteins.

Purification and Characterization of Protein Carboxyl O-Methyltransferase from Porcine Spleen

  • Yoon, Sung-Pil;Son, Min-Sik;Han, Jeung-Whan;Lee, Hyang-Woo;Hong, Sung-Youl
    • BMB Reports
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    • v.30 no.6
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    • pp.410-414
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    • 1997
  • We purified a protein carboxyl O-methyltransferase (protein methylase II) from porcine spleen to homogeneity. The molecular weight of the porcine spleen protein methylase II (ps-PM II) was estimated to be 27,500 daltons on SDS-PAGE. Amino acid sequence of N-terminal 28 residues for ps-PM II was identified. Amino-terminal three amino acid residues of ps-PM II were deleted when compared to those of other protein carboxyl methytransferase. S-Adenosyl-L-homocysteine competitively inhibits ps-PM II with a K, value of $1.63{\times}10^{-7}M$. Myelin basic protein exhibited the highest methyl-accepting capacity among the proteins tested.

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Cloning and Sequencing of Coat Protein Gene of the Korean Isolate of Rice stripe virus

  • Hong, Yeon-Kyu;Kwak, Do-Yeon;Park, Sung-Tae;Choi, Jo-Im;Lee, Key-Woon;Lee, Bong-Choon
    • The Plant Pathology Journal
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    • v.20 no.4
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    • pp.313-315
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    • 2004
  • The coat protein gene of Korean isolate of Ricer stripe virus (RSV-Kr) was cloned and its nucleotide sequence was determined. Total RNA was extracted from infected leaves and RSV viral RNA was detected by using RT-PCR with specific primer of coat protein gene. The result of RT-PCR showed a specific band. Purified RT-PCR products of coat protein gene were ligated into the pGEM-T Easy plasmid vector and cloned cDNA was obtained for nucleotide sequence determination. Coat protein gene of RSV-Kr consisted of 969 bp long encoding a protein of 322 amino acids. RSV-Kr showed 94%-99% sequence identities to that of Japanese- and Chinese isolates.

Effects of Various Reagents on Textural Properties of Soy Protein Gel (대두단백겔의 물성에 미치는 분자결합력 저해 시약의 영향)

  • 배동호;정호선
    • Food Science and Preservation
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    • v.5 no.1
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    • pp.65-71
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    • 1998
  • The changes in gel characteristics of soy protein as a result of various reagents that alter specific interactions which affect the formation and textural properties of gels, were studied. The reagents were added to 15% soy protein solutions prior to heat treatment. The gels were not formed with urea, indicating that hydrogen bonds significantly contributed to the formation and hardness of soy protein gel. Hydrophobic interactions and disulfide bonds compensated for hydrogen bonds and the contributions of electrostatic interactions to gel hardness are relatively insignificant. The farce primarily responsible for gel cohesiveness appeared to be disulfide bonds, because a significant decrease in cohesiveness was found only with the presence of N-ethylmaleimide. Adhesiveness decreased only with the addition of urea, and thus the contribution of hydrogen bonding to adhesiveness of gel could be concluded to be resent. However, adhesiveness was suggested to be interpreted not only wile molecular forces involved in gel formation but also with hydration properties of protein.

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Mutational Analysis of Cucumber Mosaic Virus Movement Protein Gene

  • You, Jin-Sam;Baik, Hyung-Suk;Paek, Kyung-Hee
    • BMB Reports
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    • v.32 no.1
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    • pp.82-85
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    • 1999
  • The movement protein of cucumber mosaic virus (CMV) is required for cell-to-cell movement of viral RNA. The movement of viral RNA occurs through the plant intercellular connection, the plasmodesmata. The viral movement protein was known to be multi-functional. In this work, a series of deletion mutants of CMV movement protein gene were created to identify the functional domains. The mutated movement proteins were produced as inclusion body in E. coli, and purified and renatured. A polyclonal antibody was raised against the CMV-Kor strain (Korean isolate) movement protein expressed in E. coli. The ability of the truncated proteins to bind to ssRNA was assayed by UV cross-linking and gel retardation analyses. The results indicate that the domain between amino acids 118 and 160 of CMV movement protein is essential for ssRNA binding.

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