• Title/Summary/Keyword: Acidic proteins

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Sequence Analysis of Hypothetical Proteins from Helicobacter pylori 26695 to Identify Potential Virulence Factors

  • Naqvi, Ahmad Abu Turab;Anjum, Farah;Khan, Faez Iqbal;Islam, Asimul;Ahmad, Faizan;Hassan, Md. Imtaiyaz
    • Genomics & Informatics
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    • v.14 no.3
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    • pp.125-135
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    • 2016
  • Helicobacter pylori is a Gram-negative bacteria that is responsible for gastritis in human. Its spiral flagellated body helps in locomotion and colonization in the host environment. It is capable of living in the highly acidic environment of the stomach with the help of acid adaptive genes. The genome of H. pylori 26695 strain contains 1,555 coding genes that encode 1,445 proteins. Out of these, 340 proteins are characterized as hypothetical proteins (HP). This study involves extensive analysis of the HPs using an established pipeline which comprises various bioinformatics tools and databases to find out probable functions of the HPs and identification of virulence factors. After extensive analysis of all the 340 HPs, we found that 104 HPs are showing characteristic similarities with the proteins with known functions. Thus, on the basis of such similarities, we assigned probable functions to 104 HPs with high confidence and precision. All the predicted HPs contain representative members of diverse functional classes of proteins such as enzymes, transporters, binding proteins, regulatory proteins, proteins involved in cellular processes and other proteins with miscellaneous functions. Therefore, we classified 104 HPs into aforementioned functional groups. During the virulence factors analysis of the HPs, we found 11 HPs are showing significant virulence. The identification of virulence proteins with the help their predicted functions may pave the way for drug target estimation and development of effective drug to counter the activity of that protein.

Effects of Auxin, GA and Cytokinin on the Protein Synthesis (Accumulation) of Soybean (Auxin, GA 및 Cytokinin이 대두의 단백질합성 (축적)에 미치는 영향)

  • Yoo, Ki-Jung;Park, Chang-Kyu;Kim, Su-Il
    • Applied Biological Chemistry
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    • v.29 no.1
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    • pp.73-77
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    • 1986
  • Aqueous solutions of 2,4-D, BA or $GA_3(10^{-6},\;10^{-5},\;and\;10^{-4}M,\;respectively)$ were sprayed onto soybean (Glycine max) plants in the flowering stage, and proteins of immature (33days after flowering) and mature (77days after flowering) seeds were analyzed by electrophoresis to elucidate the effects of the growth reguators on protein synthesis or protein accumulation in the seeds. Accumulations of some proteins were altered by 2,4-D or BA at certain concentrations, but no proteins were affected by $GA_3$. The ${\alpha}\;and\;{\alpha}'$ subunits of 7S and acidic subunit of 11S disappeared in mature seeds after treatments at the flowering stage with 2,4-D or BA. The presence of ${\alpha}\;and\;{\alpha}'$ subunits of 7S and acidic subunit of 11S in immature seeds indicated that the absence of the above polypeptides in mature seeds did not result from inhibitions in syntheses of the polypeptides by the growth regulators. Disappearance of the above proteins in mature seeds seemed to be concerned with the action of specific proteolytic enzyme (s) (metalloendopeptidase?), and 2,4-D and BA might promote gene expression or activation of the enzyme.

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pH Effect on the Structure of Reduced NifU-like Protein from Helicobacter pylori

  • Lee, Ki-Young;Kim, Ji-Hun;Bae, Ye-Ji;Lee, Bong-Jin
    • Journal of the Korean Magnetic Resonance Society
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    • v.19 no.3
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    • pp.106-111
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    • 2015
  • Helicobacter pylori (H. pylori) survives in acidic and fluctuating pH conditions of the stomach. The pH effect on H. pylori proteins is important for the advanced understanding of its evolution and viability, although this bacterium has the molecular machinery that neutralizes the acidic condition. HP1492 is known as a conserved NifU-like protein from H. pylori. NifU is a nitrogen fixation protein that mediates the transfer of iron-sulfur (Fe-S) cluster to iron-sulfur proteins like ferredoxin. Commonly, the monomeric reduced state of NifU can be converted to the dimeric oxidized state by intermolecular disulfide bond formation. Because it remains unclear that HP1492 actually behaves as known NifU protein, we first found that this protein can adopt both oxidized and reduced forms using size exclusion chromatography. Circular dichroism experiment showed that HP1492 is relatively well-structured at pH 6.5, compared to other pH conditions. On the basis of the backbone resonance assignment of HP1492, we further characterized the residues that are sensitive to pH using NMR spectroscopy. These residues showing large chemical shift changes could be mapped onto the secondary structure of the protein. Our results could provide the foundation for structural and biophysical studies on a wide spectrum of NifU proteins.

Property Comparison of Polyclonal Anti-Soy Protein Antibodies Produced for ELISA (효소면역측정법을 위한 다클론 항대두단백 항체의 생산 및 특성비교)

  • Shon, Dong-Hwa;Kim, Hyun-Jung;Yun, Sung-Seob
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1221-1226
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    • 2000
  • Specific antibodies were produced to develope the enzyme-linked immunosorbent assay for analysis of soy proteins and the properties of the antibodies were compared. Isolate soy protein(ISP), and ISP heated with SDS and urea (ISP(SU)), acidic subunits(AS) of 11S globulin were immunized to produce polyclonal antibodies. By using competitive indirect ELISA(ciELISA), the reactivities of the antibodies toward soy proteins treated with different methods were investigated and shown as $IC_{50}$. $IC_{50}'s$ of anti-ISP antibodies to ISP, ISP(SU), ISP treated with 2-ME(ISP(ME)), and crude 11S were 20, 30, 36, and $1000\;{\mu}g/mL$, respectively. And the values of anti-ISP(SU) antibodies to the same antigens were 100, 5, 4, and $220\;{\mu}g/mL$ and those of anti-AS antibodies were 20, 2, 2.5, and $200\;{\mu}g/mL$, respectively. Therefore, anti-AS antibodies showed the highest reactivities toward soy proteins among the produced antibodies as determined by ciELISA.

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Apoptotic Effect of Ethanol Extracts of Bojungbangamtang and Acidic Polysaccharide of Korea Red Ginseng in a MCF7/adR Multidrug-resistance Breast Cancer Cells (항암제 내성 유방암 MCF7/adR 세포주에 대한 보정방암탕과 홍삼산성다당체의 세포고사 유도효과)

  • Ahn, Gyu-In;Park, Cheol-Hwan;Lee, Eun-Ok;Lee, Hyo-Jung;Lee, Jae-Ho;Kim, Kwan-Hyun;Rhee, Yun-Hee;Jang, Yu-Sung;Kim, Sang-Tae;Kim, Sung-Hoon
    • YAKHAK HOEJI
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    • v.50 no.4
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    • pp.272-277
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    • 2006
  • This study was undertaken to determine whether the 9 herbal complex induces apoptosis in human breast cancer MCF-7 cells and adriamycin-resistant MCF7/adR cells. Ethanol extracts of Bojungbangamtang (BBTE) and acidic polysaccharide of Red Ginseng (GIN) induced cell death in both MCF-7 and MCF7/adR cells. Ethanol extracts of Bojungbangamtang and acidic polysaccharide of Red Ginseng also induced $G_2/M$ cell cycle arrest and increased TUNEL positive cells in MCF7/adR cells. In addition, flow cytometric analysis revealed the decreased expression of P-glycoprotein (P-gp) in ethanol extracts of Bojungbangamtang and acidic polysaccharide of Red Ginseng treated MCF7/adR cells. Similarly, decreased protein levels of P-glycoprotein and multidrug resistance associated proteins-1 were also determined by immunocytometry in ethanol extracts of Bojungbangamtang treated MCF7/adR cells. Taken together these data indicate that ethanol extracts of Bojungbangamtang and acidic polysaccharide of Red Ginseng inhibit the function of ABC transporters such as multi drug resistance associated proteins (MRPs) and P-glycoprotein as well as induce apoptosis in MCF7/adR cells. Thus, these data suggest that ethanol extracts of Bojungbangamtang and polysaccharide of Red Ginseng can be candidates for the treatment of multidrug-resistant MCF7/adR cells.

Hydrolysis of 7S and 11S Soy Proteins by Commercial Proteases (단백분해효소(蛋白分解酵素)에 의한 대두(大豆) 7S 및 11S 단백질(蛋白質)의 가수분해(加水分解))

  • Kang, Yeung-Joo;Lee, Ki-Chun;Park, Yeung-Ho
    • Korean Journal of Food Science and Technology
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    • v.20 no.3
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    • pp.338-343
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    • 1988
  • Selected kinetic parameters and degree of hydrolysis(DH) were measured using commercial proteases(trypsin, alcalase and pronase) to study the affinity of these enzymes to 7S and 11S soy proteins. Electrophoretic patterns of the hydrolysates were also investigated. In general, the order of affinity between the proteins and the proteases was 11S(protein-rich fraction)and 7S PRF for unheated proteins, and 7S PRF and 11S PRF for preheated proteins. Substrate inhibition was present at a substrate concentration of 1.5% or higher when preheated protein was used as the substrate. The maximum DH values of alcalase were obtained from 7S PRF(60%) and 11S PRF(80%) at 1 hr hydrolysis, respectively. Trypsin hydrolyses did not affect 11S soy protein but the acidic subunits in contrast to alcalase and pronase hydrolyses which changed almost all subunits. Alcalase hydrolysis induced distinct changes on 2S soy protein.

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The Scaffolding Protein WAVE1 Associates with Kinesin 1 through the Tetratricopeptide Repeat (TPR) Domain of the Kinesin Light Chain (KLC) (Kinesin Light Chain (KLC)의 Tetratricopeptide Repeat (TPR) 도메인을 통한 Scaffold 단백질 WAVE1과 Kinesin 1의 결합)

  • Jang, Won Hee;Jeong, Young Joo;Urm, Sang-Hwa;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.26 no.8
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    • pp.963-969
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    • 2016
  • Kinesin superfamily proteins (KIFs) are microtubule-dependent molecular motor proteins essential for the intracellular transport of organelles and protein complexes in cells. Kinesin 1 is a member of those KIFs that transport various cargoes, including organelles, synaptic vesicles, neurotransmitter receptors, cell signaling molecules, and mRNAs through interaction between its light chain subunit and the cargoes. Kinesin light chains (KLCs) are non-motor subunits that associate with the kinesin heavy chain (KHC) dimer. KLCs interact with many different binding proteins, but their particular binding proteins have not yet been fully identified. We used the yeast two-hybrid assay to identify proteins that interact with the tetratricopeptide repeat (TPR) domain of KLC1. We found an interaction between the TPR domain of KLC1 and Wiskott-Aldrich syndrome protein family member 1 (WAVE1), a member of the WASP/WAVE family involved in regulation of actin cytoskeleton. WAVE1 bound to the six TPR domain-containing regions of KLC1 and did not interact with KHCs (KIF5A, KIF5B, and KIF5C) in the yeast two-hybrid assay. The carboxyl (C)-terminal verprolin-cofilin-acidic (VCA) domain of WAVE1 is essential for interaction with KLC1. Also, other WAVE isoforms (WAVE2 and WAVE3) interacted with KLC1 in the yeast two-hybrid assay. When co-expressed in HEK-293T cells, WAVE1 co-localized with KLC1 and co-immunoprecipitated with KLC1 and KIF5B. These results suggest that kinesin 1 motor protein may transport WAVE complexes or WAVE-coated cargoes in cells.

Analysis of brain protein expression in developing mouse fetus (임신일령에 따른 생쥐 태아 뇌조직의 단백질 발현 양상 분석)

  • Han, Rong-Xun;Kim, Hong-Rye;Diao, Yun-Fei;Woo, Je-Seok;Jin, Dong-Il
    • Korean Journal of Agricultural Science
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    • v.38 no.1
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    • pp.65-70
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    • 2011
  • Development of mouse fetus brains can be defined morphologically and functionally by three developmental stages, embryo day (ED) 16, postnatal stage one week and eight weeks. These defined stages of brain development may be closely associated with differential gene expression rates due to limited cellular resources such as energy, space, and free water. Complex patterns of expressed genes and proteins during brain development suggests the changes in relative concentrations of proteins rather than the increase in numbers of new gene products. This study was designed to evaluate early protein expression pattern in mouse fetus brain. The mouse brain proteome of fetus at ED 15.5, and 19.5 was obtained using 2-dimensional gel electrophoresis (DE). Analysis of the 2-DE gels in pH 3-10 range revealed the presence of 15 differentially expressed spots, of which 11 spots were identified to be known proteins following MALDI-TOF analysis; 3 spots were up-regulated and 8 spots were down-regulated in the mouse fetus brain at ED 15.5. UP-regulated proteins were identified as MCG18238, isoform M2 of pyruvate kinase isozymes M1/M2, isoform 2 of heterogeneous nuclear ribonucleoprotein K, heterogeneous nuclear ribonucleoprotein H2, creatine kinase B-type, 40S ribosomal protein SA and hemoglobin subunit beta-H1. Down-regulated proteins were putative uncharacterized protein, lactoylglutathione lyase and secreted acidic cysteine rich glycoprotein. Our results revealed composite profiles of mouse fetus brain proteins related to mouse fetus development by 2-DE analysis implying possible roles of these proteins in neural differentiation.

Comparative Study of Protein Profile during Development of Mouse Placenta

  • Han, Rong-Xun;Kim, Hong-Rye;Naruse, Kenji;Choi, Su-Min;Kim, Baek-Chul;Park, Chang-Sik;Jin, Dong-Il
    • Reproductive and Developmental Biology
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    • v.31 no.4
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    • pp.253-260
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    • 2007
  • To examine the differential protein expression pattern in the 11.5 day post-coitus (dpc) and 18.5 dpc placenta of mouse, we have used the global proteomics approach by 2-D gel electrophoresis (2-DE) and MALDI-TOF-MS. The differential protein patterns of 3 placentae at the 11.5 dpc and 18.5 dpc from nature mating mice were analyzed. Proteins within isoelectric point range of $3.0{\sim}10.0$, separately were analyzed in 2DE with 3 replications of each sample. A total of approximately 1,600 spots were detected in placental 2-D gel stained with Coomassie-blue. In the comparison of 11.5 dpc and 18.5 dpc placentae, a total of 108 spots were identified as differentially expressed proteins, of which 51 spots were up-regulated proteins such as alpha-fetoprotein, mKIAA0635 protein and transferrin, annexin A5, while 48 spots were down-regulated proteins such as Pre-B-cell colony-enhancing factor l(PBEF), aldolase 1, A isoform, while 4 spots were 11.5 dpc specific proteins such as chaperonin and Acidic ribosomal phosphoprotein P0, while 3 spots were 18.5 dpc specific proteins such as aldo-keto reductase family 1, member B7 and CAST1/ERC2 splicing variant-1. Most identified proteins in this analysis appeared to be related with catabolism, cell growth, metabolism and regulation. Our results revealed composite profiles of key proteins involved in mouse placenta during pregnancy.

Proteome Analysis of the Young Spikelets of Photoperiod-Sensitive Rice Mutant Treated in Different Photoperiods

  • Pandeya, Devendra;Song, You-Chun;Kim, Sung-Su;Suh, Hak-Soo;Kang, Sang-Gu
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.52 no.3
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    • pp.281-288
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    • 2007
  • Photoperiod sensitive genetic male sterile (PGMS) rice is sterile mutant controlled by photoperiod. A PGMS mutant 920S was sterile grown under long-day (LD) photoperiod (14 h light/10 h dark) but fertile grown under short-day (SD) photoperiod (10 h light/14 h dark). Proteome analysis revealed that 12 protein spots were differentially expressed in the spikelets of 920S plants either treated with LD or SD photoperiod. Among these proteins, three proteins including chlorophyll a/b binding protein, vacuolar ATPase ${\beta}-subunit,\;{\alpha}-tubulin$ and an unknown protein were more than three-fold abundant in the spikelet of the SD-treated plants than those of the LD-treated plants. On the other hand, eight proteins including acetyl transferase, 2, 3- biphosphoglycerate, aminopeptidase N, pyruvate decarboxylase, 60S acidic ribosomal protein and three unknown protein spots were more abundant in the spikelets of the LD-treated plants than those of the SD-treated plants. The results suggest that the observed proteins may be involved in sterile or fertile pollen development under LD or SD photoperiod respectively in the PGMS mutant rice.