• Title/Summary/Keyword: protein binding

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Cis-acting Elements in the 3' Region of Potato virus X are Required for Host Protein Binding

  • Kwon, Sun-Jung;Kim, Kook-Hyung;Hemenway Cynthia
    • The Plant Pathology Journal
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    • v.22 no.2
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    • pp.139-146
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    • 2006
  • The 3' region of Potato virus X (PVX) has the 74 nt 3'-nontranslated region (NTR) that is conserved among all potexviruses and contains several cis-acting elements for minus-strand and plus-strand RNA accumulation. Three stem-loop structures (SL1-SL3), especially formation of SL3 and U-rich sequence of SL2, and near upstream elements in the 3' NTR were previously demonstrated as important cis-acting elements. To Investigate the binding of these cis-acting elements within 3' end with host protein, we used the electrophoretic mobility shift assays (EMSA) and UV-cross linking analysis. The EMSA with cellular extracts from tobacco and RNA transcripts corresponding to the 150 nt of the 3' end of PVX RNA showed that the 3' end of PVX formed complexes with cellular proteins. The specificity of protein binding was confirmed through competition assay by using with 50-fold excess of specific and non-specific probes. We also conducted EMSA with RNAs containing various mutants on those cis-acting elements (${\Delta}10$10, SL3B, SL2A and ${\Delta}21$; J Mol Biol 326, 701-720) required for efficient PVX RNA accumulation. These analyses supported that these cis-acting elements are required for interaction with host protein(s). UV-cross linking analysis revealed that at least three major host proteins of about 28, 32, and 42 kDa in mass bound to these cis-elements. These results indicate that cis-acting elements from 3' end which are important for minus and plus-strand RNA accumulation are also required for host protein binding.

Molecular cloning and nucleotide sequence of schizosaccharomyces pombe Homologue of the receptor for activated protein kinase C gene

  • Park, Seung-Keil;Yoo, Hyang-Sook
    • Journal of Microbiology
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    • v.33 no.2
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    • pp.128-131
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    • 1995
  • Using differential hybridization, we selected the prk gene fortuitously from Schizosaccharomyces pombe homologous to RACK1 of rat which encodes the receptor for activated protein kinase C. The cDNA sequence of prk was determined and its deduced amino acid sequence was 76% homologous to RACK1 and had the feature of trimeric G protein bata subunit. The specific amino acid sequences required for the protein kinase C binding were also present in Prk as in the case of RACK1 protein. From these similarities, we suggest that the Prk is protein kinase C binding protein of S. prombe. The involvement of Prk in signal transduction mediated by protein kinase C remained to be studied.

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The Search of Pig Pheromonal Odorants for Biostimulation Control System Technologies: Ⅱ. Holographic QSAR Model for Binding Affinities between Ligands of Volatile Odorants Molecules and Porcine Odorant Binding Protein (pOBP) (생물학적 자극 통제 수단으로 활용하기 위한 돼지 페로몬성 냄새 물질의 탐색: Ⅱ. 휘발성 냄새분자의 리간드와 Porcine Odorant Binding Protein (pOBP) 사이의 결합 친화력에 관한 홀로그래피적 QSAR 모델)

  • Sung N. D.;Park C. S.;Choi Y. S.;Myung P. K.
    • Reproductive and Developmental Biology
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    • v.29 no.1
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    • pp.43-48
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    • 2005
  • To search of a new porcine pheromonal odorants for biostimulation control system technologies to offer a potentially useful and practical way to improve reproductive efficiency in livestock species, the holographic quantitative structure activity relationship (HQSAR) model between odorants, 2-phenoxytetrahydrofurane (A), 2-cyclohexyl-oxytetrahydrofurane (B), derivatives and binding affinity constants (p[Od.]/sub 50/) for porcine odorant-binding protein (pOBP) as receptor of pig pheromones were derivated and disscused. The binding affinity constants of cyclohexyl substituents (A) for pOBP were higher (A>B) than that of phenyl substituents (B). It was revealed that the optimum HQSAR model XI using PLS analyses had a fragment length (5∼8) with chirality at 5 components and hologram length 97 bin, which had a cross-validated q²(predictablities) of 0.916, and a conventional correlation coefficient r² (fitness) of 0.988, respectively. From the atomic contribution, the C3 and C5 atom in 2-oxyfuryl group contributed to binding affinity constants, whereas the central carbon atom in tert-butyl group on the cyclohexyl ring and the C4 atom of furyl group parts showed no contribution.

The In Vitro Translocation of Escherichia coli Ribose-binding Protein via Various Targeting Routes

  • Lee, Byoung-Chul;Kim, Hyoung-Nan;Hwang, Yong-Il
    • BMB Reports
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    • v.34 no.2
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    • pp.118-122
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    • 2001
  • The translocation of ribose-binding protein (RBP) into the inverted membrane vesicles (IMV) of Escherichia coli and eukaryotic microsomes was studied using the in vitro translation/translocation system. It was found that RBP was translocated into heterologous eukaryotic microsomes co-translationally, as well as post-translationally However, RBP was translocated only past-translationally into IMV. Degradation fragments of RBP with the molar mass of 14 and 16 kDa were produced during the translocation into IMV However, the amount of the degradation products decreased and the mature form of RBP appeared in the presence of phenylmethylsulfonyl fluoride (PMSF). PMSF and GTP accelerated the translocation of RBF It was also found that SecB enhanced the post-translational translocation of RBP It appears that RBP is translocated via at least two targeting paths.

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Stability and Structural Change of cAMP Receptor Protein at Low and High cAMP Concentrations

  • GANG JONGBACK;CHUNG HYE-JIN;PARK GWI-GUN;PARK YOUNG-SEO;CHOI SEONG-JUN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1392-1396
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    • 2005
  • Proteolytic digestion and CD measurement of wild-type and mutant cyclic AMP receptor proteins (CRPs) were performed either in the presence or absence of cyclic nucleotide. Results indicated that transition of a structural change to the hinge region by the binding of cAMP to the anti site was required for the binding of cAMP to the syn site near the hinge region and, although the occupancy of cAMP in the anti site increased the protein stability, CRP adopted more a stable conformation by the binding of cAMP to the syn site.

Identification of Ran-binding protein M as a stanniocalcin 2 interacting protein and implications for androgen receptor activity

  • Shin, Jihye;Sohn, Young Chang
    • BMB Reports
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    • v.47 no.11
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    • pp.643-648
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    • 2014
  • Stanniocalcin (STC), a glycoprotein hormone originally discovered in fish, has been implicated in calcium and phosphate homeostasis. While fishes and mammals possess two STC homologs (STC1 and STC2), the physiological roles of STC2 are largely unknown compared with those of STC1. In this study, we identified Ran-binding protein M (RanBPM) as a novel binding partner of STC2 using yeast two-hybrid screening. The interaction between STC2 and RanBPM was confirmed in mammalian cells by immunoprecipitation. STC2 enhanced the RanBPM-mediated transactivation of liganded androgen receptor (AR), but not thyroid receptor ${\beta}$, glucocorticoid receptor, or estrogen receptor ${\beta}$. We also found that AR interacted with RanBPM in both the absence and presence of testosterone (T). Furthermore, we discovered that STC2 recruits RanBPM/AR complex in T-dependent manner. Taken together, our findings suggest that STC2 is a novel RanBPM-interacting protein that promotes AR transactivation.

Hemolymph Juvenile Hormone Binding Protein of Fifth Instar Larvae of Bombyx mori L.: Identification and Purification (누에나방의 5령유충 혈림프의 유약호르몬 결합단백질: 확인 및 정제)

  • Park, Chul-Ho;Kim, Hak-Ryul
    • The Korean Journal of Zoology
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    • v.37 no.1
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    • pp.66-75
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    • 1994
  • Juvenile hormone binding protein was identified in the hemolymph of fifth instar larvae and purified using column chromatography. Hemolymph was mixed with [3H] JH-III and electrophoresed on 691 NON-SDS gel, indicating that radioactivity peak appears at Rf value of 0.55. Gel filtration showed two radioactivity peaks equivalent to bound and free [3H]JH-III, respectively. JHBP was purified from hemolymph through gel filtration (Sephadex G-100), anion exchange chromatosraphv (DEAE Sepharose CL-6B), chromatofocusing chromatographv (PBE 94) and preparative electrophoresis.

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Regulation of Cyclic AMP-Response Element Binding Protein Zhangfei (CREBZF) Expression by Estrogen in Mouse Uterus

  • Jang, Hoon
    • Development and Reproduction
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    • v.22 no.1
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    • pp.95-104
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    • 2018
  • CREBZF (cAMP-response element binding protein zhangfei) is a member of ATF/CREB family, and which regulates various cellular functions by suppressing major factors with direct interaction. In this study, we have examined the expression of CREBZF on mouse endometrium during uterus estrous cycles and estrogen (E2) treatment. In uterus, CREBZF mRNA expression was higher than other organs and mRNA and protein of CREBZF was increased in proestrus phase and decreased in estrus phase. The expression of CREBZF in 3-weeks old mouse uterus was reduced by E2 injection in endometrium. In addition, the expression of progesterone receptor, a marker of E2 in ovariectomized mice was found to be strongly expressed in stroma, while CREBZF was only expressed in epithelium. Also, we conformed that E2-suppressed CREBZF was restored by co-injection of ICI 182,780, an estrogen receptor antagonist. Overall, these results suggest that CREBZF is regulated by estrogen and involved in ER signaling pathway in mouse uterus.

Binding sites for lead ion in staphylococcus epidermidis

  • Kim, Mal-Nam;Sung, Hye-Yoon
    • Journal of Microbiology
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    • v.33 no.3
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    • pp.228-233
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    • 1995
  • As S. epidermidis cell was fractionated into cell wall, cell membrane, and cytoplasm, the cell membrane proved to be the most efficient absorbent for lead ion. Utrasonication was effective, when the cells were treated during their exponential growth. The amount of the lead ion adsorbed in cell membrane decreased as hydrogen ion concentration of solution increased. Protein purified from the cell membrane showed higher adsorption capacity for the lead ion than peptidoglycan, teichoic acid from cell wall, or cell membrane lipid. Modification of carboxyl groups in the membrane protein with ethylenediamine and 1-ethyl-3-carbodiimide hydrochloride resulted in a considerable decrease of lead ion adsorption capability, suggesting that the main binding site for lead ion was the carboxyl groups of protein in cell membrane.

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The Search of Pig Pheromonal Odorants for Biostimulation Control System Technologies: A 2D-QSAR Model for Binding Affinity between 2-Cyclohexyloxytetrahydrofurane Analogues and Porcine Odorant Binding Protein (생물학적 자극 통제 수단으로 활용하기 위한 돼지 페로몬성 냄새 물질의 탐색: 2-Cyclohexyloxytetrahydrofurane 유도체와 Porcine Odorant Binding Protein 사이의 결합 친화력에 관한 2D-QSAR 모델)

  • Park, Chang-Sik;Choi, Yang-Seok;Sung, Nack-Do
    • Reproductive and Developmental Biology
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    • v.31 no.1
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    • pp.15-20
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    • 2007
  • To search of a new porcine pheromonal odorant for biostimulation control system technologies to offer a potentially useful and practical way to improve reproductive efficiency in livestock species, the two dimensional quantitative structure-activity relationship (QSAR) models between physicochemical parameters as descriptors of 2-cyclohexyloxytetrahydrofurane (A), 2-phenoxytetrahydrofurane (B) analogues and binding affinity constant ($p[Od.]_{50}$) for porcine odorant-binding protein (pOBP) as receptor of pig pheromones were derived and disscused. The statistical quality of the optimized 2D-QSAR model is good ($r^{2}=0.964$) and accounts for 96.4% of the variance in the binding affinity constants. It was found that the binding affinity constants were dependent upon the optimal value, $(SL)_{opt.}=1.418$ of substituent lipole (SL) in molecules. Therefore, the SL constant was very important factor for binding affinity.