• 제목/요약/키워드: binding proteins

검색결과 1,465건 처리시간 0.026초

Binding Characteristics to Mosquito-larval Midgut Proteins of the Cloned Domain II-III Fragment from the Bacillus thuringiensis Cry4Ba Toxin

  • Moonsom, Seangdeun;Chaisri, Urai;Kasinrerk, Watchara;Angsuthanasombat, Chanan
    • BMB Reports
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    • 제40권5호
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    • pp.783-790
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    • 2007
  • Receptor binding plays an important role in determining host specificity of the Bacillus thuringiensis Cry $\delta$-endotoxins. Mutations in domains II and III have suggested the participation of certain residues in receptor recognition and insect specificity. In the present study, we expressed the cloned domain II-III fragment of Cry4Ba and examined its binding characteristics to mosquito-larval midgut proteins. The 43-kDa Cry4Ba-domain II-III protein over-expressed in Escherichia coli as inclusion bodies was only soluble when carbonate buffer, pH 10.0 was supplemented with 4M urea. After renaturation via stepwise dialysis and subsequent purification, the refolded domain II-III protein, which specifically reacts with anti Cry4Ba-domain III monoclonal antibody, predominantly exists as a $\beta$-sheet structure determined by circular dichroism spectroscopy. In vitro binding analysis to both histological midgut tissue sections and brush border membrane proteins prepared from susceptible Aedes aegypti mosquito-larvae revealed that the isolated Cry4Ba-domain II-III protein showed binding functionality comparable to the 65-kDa full-length active toxin. Altogether, the data present the 43-kDa Cry4Ba fragment comprising domains II and III that was produced in isolation was able to retain its receptor-binding characteristics to the target larval midgut proteins.

Identification and Functional Analysis of SEDL-binding and Homologue Proteins by Immobilized GST Fusion and Motif Based Methods

  • Hong, Ji-Man;Jeong, Mi-Suk;Kim, Jae-Ho;Kim, Boog-il;Holbrook, Stephen R.;Jang, Se-Bok
    • Bulletin of the Korean Chemical Society
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    • 제29권2호
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    • pp.381-388
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    • 2008
  • An X-linked skeletal disorder, SEDT (spondyloepiphyseal dysplasia tarda) is a genetic disease characterized by a disproportionately short trunk and short stature caused by mutations in the SEDL gene. This gene is evolutionarily conserved from yeast to human. The yeast SEDL protein ortholog, Trs20p, has been isolated as a member of a large multi-protein complex called the transport protein particle (TRAPP), which is involved in endoplasmic reticulum (ER)-to-Golgi transport. The interaction between SEDL and partner proteins is important in order to understand the molecular mechanism of SEDL functions. We isolated several SEDL-binding proteins derived from rat cells by an immobilized GST-fusion method. Furthermore, the SEDL-homologue proteins were identified using motif based methods. Common motifs between SEDL-binding proteins and SEDL-homologue proteins were classified into seven types and 78 common motifs were revealed. Sequence similarities were contracted to seven types using phylogenetic trees. In general, types I-III and VI were classified as having the function of acetyl-CoA carboxylase, glycogen phosphorylase, isocitrate dehydrogenase, and enolase, respectively, and type IV was found to be functionally related to the GST protein. Types V and VII were found to contribute to TRAPP vesicle trafficking.

Identification of a Protein that Interacts with Calcium-Binding Protein 3(CBP3) in Dictyostelium discoideum

  • Jung, Sun-Young;Lee, Chang-Hun;Kang, Sa-Ouk
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 2001년도 학술 발표회 진행표 및 논문초록
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    • pp.43-43
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    • 2001
  • In cells of the eukaryotic microorganism Dictyostelium discoideum, at least eight small, four-EF hand calcium-binding proteins respectively are expressed at specific stages during development. One of these proteins, calcium-binding protein 3 (CBP3), first appears just prior to cell aggregation and then maintains relatively constant levels throughout development.(omitted)

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카드뮴 결합 고분자량 단백질의 생화학적 특성 (Biochemical Characteristics of Cd-binding High Molecular Weight Proteins)

  • 천기정;김봉희
    • 약학회지
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    • 제39권4호
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    • pp.380-384
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    • 1995
  • The isoelelectric points of Cd-BP(l) and Cd-BP(II), cadmium-binding proteins, were 6.01 and 5.35, respectively. Both of them contained zinc. As for the amino acid composition, Cd-BP(I) contained a lot of glycine and lysine but none of such aromatic amino acids as tyrosine and phenylalanine.. On the other hand, Cd-BP(II) contained leucine, histidine, asparti cacid and alanine but no aromatic amino acids.

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PLP-1 Binds Nematode Double-stranded Telomeric DNA

  • Im, Seol Hee;Lee, Junho
    • Molecules and Cells
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    • 제20권2호
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    • pp.297-302
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    • 2005
  • The integrity and proper functioning of telomeres require association of telomeric DNA sequences with specific binding proteins. We have characterized PLP-1, a $PUR{\alpha}$ homolog encoded by F45E4.2, which we previously identified as a candidate double stranded telomere binding protein, by affinity chromatography followed by mass spectrometry. PLP-1 bound double-stranded telomeric DNA in vitro as shown by competition assays. Core binding was provided by the third and fourth nucleotides of the TTAGGC telomeric repeat. This is quite different from the binding sequence of CEH-37, another C. elegans telomere binding protein, suggesting that multiple proteins may bind nematode telomeric DNA simultaneously in vivo.

STRUCTURAL ANALYSIS OF RAPAMYCIN'S ROLE IN BINDING FKBP12 AND FRAP

  • Park, Jungwon;Jie Chen;Stuart Schreiber;Jon Clardy
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 1996년도 정기총회 및 학술발표회
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    • pp.9-9
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    • 1996
  • The immunosuppressive and cell cycle arrest agent rapamycin works by binding together two proteins: the FK506 binding protein (FKBP12) and the FKBP-rapamycin associated protein (FRAP). A 2.7 $\AA$ resolution crystal structure of the triple complex of human FK506 binding protein (FKBP12), rapamycin, and FKBP12-rapamycin binding domain (FRB) of FRAP, reveals two proteins bound together through rapamycin' s ability to simultaneously occupy two different hydrophobic binding pockets. (omitted)

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Binding of IciA protein to the dnaA promoter region

  • Kim, Hakjung;Hwang, Deog-Su
    • Journal of Microbiology
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    • 제33권3호
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    • pp.191-195
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    • 1995
  • IciA protein has been shown as an inhibitor for the initiation of E. coli chromosomal DNA replication at oriC. IciA protein binds the AT-rich region in oriC and then blocks the initiation of chromosomal DNA replication. Two binding sites for IciA protein were identified in dnaA gene, encoding the initiator for the E. coli chromosomal replication, promoter region by gel-shift assay and DNase I footprinting, One, named as IciA site I, is located upstream of the dnaA promoter 1P. The other, named as IciA site II, is located downstream of the dnaA promoter 2P. The sequence comparison of the regions protected from the DNase I cleavage did not result in a clear consensus sequence for the binding of IciA protein, suggesting that IciA protein may be a member of multimeric complex dsDNA binding proteins. This study provided information about the binding mode of IciA protein. Even though the IciA site II and IciA binding site in oriC seem to be composed of two IciA binding units, one binding unit is likely enough to cause the binding of IciA protein to the IciA site I. The binding of IciA protein to the dna4 promoter implies that IciA protein may involve not only the control of the initiation of chromosomal DNA replication but also the control of the dna4 gene expression.

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The Alpha Subunit of Go Interacts with Promyelocytic Leukemia Zinc Finger Protein

  • Ghil Sung-Ho
    • 대한의생명과학회지
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    • 제10권4호
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    • pp.407-413
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    • 2004
  • Heterotrimeric GTP binding proteins (G proteins) transduce signals of a variety of hormones and neurotransmitters. Go is one of the most abundant G proteins in the brain and classified as the Gi/Go family due to their sequence homology to Gi proteins. While the Gi proteins inhibit adenylyl cyclase and decrease the intracellular cAMP concentration, the functions of Go is not clearly understood despite their sequence homology to Gi. The promeylocytic leukemia zinc finger protein (PLZF) is a DNA binding transcription factor and is expressed highly in central nervous system (CNS). Several studies reported that PLZF may be involved in regulation segmentation/differentiation during CNS development. Here, I report that the alpha subunit of Go (Go ) interacts with PLZF. The interaction between Goa and PLZF was verified by using GST pulldown assay and co-immunoprecipitation. Our findings indicate that Goa could modulate gene expression via interaction with PLZF during neuronal or brain development.

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Immunoreactivity of Calcium-Binding Proteins in the Central Auditory Nervous System of Aged Rats

  • Hong, Seok-Min;Chung, Seung-Young;Park, Moon-Sun;Huh, Young-Buhm;Park, Moon-Suh;Yeo, Seung-Gun
    • Journal of Korean Neurosurgical Society
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    • 제45권4호
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    • pp.231-235
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    • 2009
  • Objective : While many factors contribute to aging, changes in calcium homeostasis and calcium related neuronal processes are likely to be important. High intracellular calcium is toxic to cells and alterations in calcium homeostasis are associated with changes in calcium-binding proteins, which confine free $Ca^{2+}$. We therefore assayed the expression of the calcium binding proteins calretinin and calbindin in the central auditory nervous system of rats. Methods : Using antibodies to calretinin and calbindin, we assayed their expression in the cochlear nucleus, superior olivary nucleus, inferior colliculus, medial geniculate body and auditory cortex of young (4 months old) and aged (24 months old) rats. Results : Calretinin and calbindin staining intensity in neurons of the cochlear nucleus was significantly higher in aged than in young rats (p<0.05) The number and staining intensity of calretinin-positive neurons in the inferior colliculus, and of calbindin-positive neurons in the superior olivary nucleus were greater in aged than in young rats (p<0.05). Conclusion : These results suggest that auditory processing is altered during aging, which may be due to increased intracellular $Ca^{2+}$ concentration, consequently leading to increased immunoreactivity toward calcium-binding proteins.

일본흰개미 (Reticulitermes speratus)의 Odorant-Binding Proteins (OBPs) 발현 양상 (Expression Patterns of Odorant-Binding Proteins (OBPs) in a Termite (Reticulitermes speratus))

  • 나종범;김재경;김규혁
    • Journal of the Korean Wood Science and Technology
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    • 제33권1호통권129호
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    • pp.1-6
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    • 2005
  • 본 연구는 국내에 서식하는 일본흰개미(Reticulitermes speratus)의 생리적 특성에 관한 기초연구로서 흰개미의 커뮤니케이션 수단인 페로몬을 후각수용기로 이동시키는 odorant-binding proteins (OBPs)의 발현 양상을 조사하기 위해서 수행되었다. 경남 진주에서 채집된 일개미와 병정개미로부터 더듬이와 다리를 각각 떼어낸 후 OBP가 존재하는지 확인하기 위하여 RT-PCR을 수행하였다. 일개미의 경우 OBP-1의 발현은 더듬이와 다리에서 모두 관찰되었으며, OBP-2와 OBP-3의 발현은 오직 더듬이에서만 관찰할 수 있었다. 병정개미의 경우 OBP-1의 발현은 일개미에서와 마찬가지로 더듬이와 다리 모두에서 관찰할 수 있었으나, OBP-2가 관찰되지 않았으며, 일개미의 더듬이에서만 관찰되었던 OBP-3의 발현을 병정개미에서는 더듬이와 다리 모두에서 볼 수 있었다. 일개미와 병정개미에서 OBP 발현 양상의 차이는 그들의 역할에 따른 분화적 특이성에 기인한 것으로 생각된다.