• Title/Summary/Keyword: crystal proteins

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Characterization and Identification of Bacillus thuringiensis subsp. tenebrionis SR6 and SR8

  • Kim, Il-Gi;Lee, Jae-Wook;Suh, Suk-Chul;Rhim, Seong-Lyul
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.772-776
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    • 2004
  • Physiological and molecular characteristics of Bacillus thuringiensis SR6 and SR8 were investigated, and phase contrast and electron microscopies revealed that a large rhomboidal crystal protein was present in the sporulating cells. SDS-PAGE and Western blot analyses showed that B. thuringiensis SR8 produced 70 kDa protein much more than other proteins, and that the 70 kDa protein could bind to the antibody of B. thuringiensis subsp. tenebrionis-crystal toxin protein, indicating that the crystal 70 kDa protein has an immunological homology with B. thuringiensis subsp. tenebrionis-crystal toxin protein. The DNA fragment of B. thuringiensis subsp. tenebrionis-toxin gene was detected in B. thuringiensis SR6 and SR8 by using PCR amplification analysis. Furthermore, the insect bioassay showed the insecticidal activity against Colorado potato beetle larvae. Based on the physiological and molecular similarities to B. thuringiensis subsp. tenebrionis, it is suggested that the B. thuringiensis SR6 and SR8 may be mutants of the B. thuringiensis subsp. tenebrionis strain overexpressing the crystal of 70 kDa toxin protein.

Isolation and Characterization of Citrobacter sp. Mutants Defective in Decolorization of Crystal Violet (Crystal vilet 색소분해능이 소실된 Citrobacter sp. 의 분리 및 특성)

  • Kim, Ji-Yoon;Kim, Kyung-Woon;Park, Yong-Lark;Cho, Young-Su;Lee, Young-Choon
    • Journal of Life Science
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    • v.10 no.4
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    • pp.333-339
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    • 2000
  • To identify genes involved in the decolorization of crystal violet, we isolated random mutants generated by transponson insertion in crystal violet-declorizing bacterium, Citrobacter sp. The resulting mutant bank yielded mutants with six distinct phenotypes, and Southern hybridization with a Tn5 fragment as a probe showed a single hybridized with six distinct phenotypes, and Southern hybridization with a Tn5 fragment as a probe showed a single hybridized band in the mutants Ctg 2, 5 an 6, whereas two and three bands were detected in Ctg1, 4 and 3, respectively. Tn5-inserted genes were isolated and the DNA sequence flanking Tn5 was determined. From comparison with a sequence database, putative protein product encoded by ctg 5 was identified as E. coli maltose transproter(Mal G) homolog, whereas the deduced amino acid sequence of the other ctg genes did not show any significant similarity with any DNA or protein sequency. Therefore, these results indicate that the other ctg genes except ctg 5 encode new proteins responsible for decolorization of crystal violet.

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Strategies in Protein Immobilization on a Gold Surface

  • Park, Jeho;Kim, Moonil
    • Applied Science and Convergence Technology
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    • v.24 no.1
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    • pp.1-8
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    • 2015
  • Protein immobilization on a gold surface plays an important role in the usefulness of biosensors that utilize gold-coated surfaces such as surface plasmon resonance (SPR), quartz crystal microbalance (QCM), etc. For developing high performance biosensors, it is necessarily required that immobilized proteins must remain biologically active. Loss of protein activity and maintenance of its stability on transducer surfaces is directly associated with the choice of immobilization methods, affecting protein-protein interactions. During the past decade, a variety of strategies have been extensively developed for the effective immobilization of proteins in terms of the orientation, density, and stability of immobilized proteins on analytical devices operating on different principles. In this review, recent advances and novel strategies in protein immobilization technologies developed for biosensors are briefly discussed, thereby providing an useful information for the selection of appropriate immobilization approach.

Validation on the molecular docking efficiency of lipocalin family of proteins

  • Sokalingam, Sriram;Munussami, Ganapathiraman;Kim, Jung-Rae;Lee, Sun-Gu
    • Journal of Industrial and Engineering Chemistry
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    • v.67
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    • pp.293-300
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    • 2018
  • Lipocalins are diverse group of small extracellular proteins found in various organisms. In this study, members of 10 non-homologous lipocalin-ligand crystal complex structures were remodeled using rigid and flexible ligand modes to validate the prediction efficiency of molecular docking simulation. The modeled ligand conformations indicated a high prediction accuracy in rigid ligand mode using cluster based analysis for most cases whereas the flexible ligand mode required further considerations such as ligand binding energy and RMSD for some cases. This in silico study is expected to serve as a platform in the screening of novel ligands against lipocalin family of proteins.

Expression of Bacillus thuringiensis CrylAc Protein Fused with Coat Protein of Potato Leafroll Virus

  • Park, Jae-Young;Li, Ming-Shun;Roh, Jong-Yul;Shim, Hee-Jin;Park, Beom-Seok;Je, Yeon-Ho
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.10a
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    • pp.116-117
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    • 2003
  • Aphidicidal activity of Bacillus thurigiensis crystal proteins was recently reported. However, relatively higher dose of crystal protein was needed to kill aphids. In this study, we intended to improve the aphidicidal activity of crystal protein by fusion with coat protein (CP) of potato leafroll virus (PLRV) which is transmitted by aphids. (omitted)

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Characterization of Peptide Deformylase2 from B. cereus

  • Park, Joon-Kyu;Kim, Kook-Han;Moon, Jin-Ho;Kim, Eunice Eun-Kyeong
    • BMB Reports
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    • v.40 no.6
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    • pp.1050-1057
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    • 2007
  • Peptide deformylase (PDF) is a metalloenzyme that removes the N-terminal formyl groups from newly synthesized proteins. It is essential for bacterial survival, and is therefore-considered as a potential target for antimicrobial chemotherapy. However, some bacteria including medically relevant pathogens possess two or more def-like genes. Here we have examined two PDFs from Bacillus cereus. The two share only 32% sequence identity and the crystal structures show overall similarity with PDF2 having a longer C-terminus. However, there are differences at the two active sites, and these differences appear to contribute to the activity difference seen between the two. BcPDF2 is found as a dimer in the crystal form with two additional actinonin bound at that interface.

Characterization of Two Type Crystal Proteins Produced by Transformed Bacillus thuringiensis NT0423 (형질전환된 Bacillus thuringiensis NT0423 균주에 의해 생산된 두가지 형태의 내독소 단백질의 특성)

  • 박현우;김호산;유용만;김상현;서숙재;강석권
    • Microbiology and Biotechnology Letters
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    • v.21 no.5
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    • pp.428-434
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    • 1993
  • Cloning and expression of two different crystal protein genes from transformed Bacillus thuringiensis were investigated. B. thuringiensis NT0423 is toxic to both Lepidopterous and Dipte-rous larvae. The pCG5 vector carrying crystal protein genes (mosquitocidal and hemolytic activity) of B. thurigiensis subsp. morrisoni PG-14 was transformed into B. thurigiensis NT0423. Transformant has expressed two type crystals of bipyramid from NT0423 and ovoid from pCG5 in one cell.

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Immunological Analysis of Antigenic Variation of Bacillus thuringiensis subsp. sotto during Sporulation and Crystallization

  • Cho, Jae Min;Gi Bum Nam;Soon Bok Hong;Myung Hwan Cho
    • Journal of Microbiology and Biotechnology
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    • v.5 no.6
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    • pp.359-363
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    • 1995
  • The antigenic variation of B. thuringiensis subsp. satto have been investigated for 120 hours during sporulation and crystallization by using SDS-PAGE and Western blot. Most antigens of a vegetative cell were found to disappear as it was in sporulation and crystallization, but protein antigens of 46, 29, 27, and 21 kDa continued to be expressed. The new protein bands of 293, 138, 119, 75, and 68 kDa appeared on days 2 through 5 in modified GYS medium. They were thought to be involved in sporulation and crystallization. The protein of 138 kDa was found to be a major protein of both crystal and spore. The expression patterns were immunologically analyzed by Western blot. The polyclonal antisera against the intact crystal showed strong immunoreactivity to proteins with molecular masses of 293, 138, 68, and 46 kDa. The polyclonal antisera against the spore recognized proteins of 293, 138, 68, and 46 kDa. Both crystals and spores appeared to express the common protein antigens.

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Degradation of the Parasporal Crystal Proteins of Bacillus thuringiensis and Induction of Stress Protein Synthesis in Bt $\delta$-endotoxin Ingested Larvae of Fall Webworm, Hyphantria cunea (Bacillus thuringiensis 의 내독소 단백질의 분해와 흰불나방 섭취유충에서 스트레스 단백질 합성의 유발)

  • 전향미;유용만;강석권;서숙재
    • Korean journal of applied entomology
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    • v.33 no.3
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    • pp.178-183
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    • 1994
  • The prolein components of each crystal toxin of Bacillus thuringiensis kurstaki and alzawai were separated by SDS-PAGE. The major clqistal proteins from both shins were composed of paiypeptides having molecular weights of 130 kd and 64 kd. Digestive mixture of both. toxins with t~ypsin and gut juices shared 62 kd polypeptide which may be major actwe toxh. However, aizawal produced much less amount of 62 kd polypeptide than kurstaki did. On ingestion with Bt &-endotoxin, larvae of Hyphantria cunea developed 45 kd stress protein in the several tissues including fat body, which was induced by heat and cold shock.

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Oligomer Model of PB1 Domain of p62/SQSTM1 Based on Crystal Structure of Homo-Dimer and Calculation of Helical Characteristics

  • Lim, Dahwan;Lee, Hye Seon;Ku, Bonsu;Shin, Ho-Chul;Kim, Seung Jun
    • Molecules and Cells
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    • v.42 no.10
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    • pp.729-738
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    • 2019
  • Autophagy is an important process for protein recycling. Oligomerization of p62/SQSTM1 is an essential step in this process and is achieved in two steps. Phox and Bem1p (PB1) domains can oligomerize through both basic and acidic surfaces in each molecule. The ZZ-type zinc finger (ZZ) domain binds to target proteins and promotes higher-oligomerization of p62. This mechanism is an important step in routing target proteins to the autophagosome. Here, we determined the crystal structure of the PB1 homo-dimer and modeled the p62 PB1 oligomers. These oligomer models were represented by a cylindrical helix and were compared with the previously determined electron microscopic map of a PB1 oligomer. To accurately compare, we mathematically calculated the lead length and radius of the helical oligomers. Our PB1 oligomer model fits the electron microscopy map and is both bendable and stretchable as a flexible helical filament.