• Title/Summary/Keyword: molecular precursor

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Improvement of protein identification performance by reinterpreting the precursor ion mass tolerance of mass spectrum (질량스펙트럼의 펩타이드 분자량 오차범위 재해석에 의한 단백질 동정의 성능 향상)

  • Gwon, Gyeong-Hun;Kim, Jin-Yeong;Park, Geon-Uk;Lee, Jeong-Hwa;Baek, Yung-Gi;Yu, Jong-Sin
    • Bioinformatics and Biosystems
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    • v.1 no.2
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    • pp.109-114
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    • 2006
  • In proteomics research, proteins are digested into peptides by an enzyme and in mass spectrometer, these peptides break into fragment ions to generate tandem mass spectra. The tandem mass spectral data obtained from the mass spectrometer consists of the molecular weights of the precursor ion and fragment ions. The precursor ion mass of tandem mass spectrum is the first value that is fetched to sort the candidate peptides in the database search. We look far the peptide sequences whose molecular weight matches with precursor ion mass of the mass spectrum. Then, we choose one peptide sequence that shows the best match with fragment ions information. The precursor ion mass of the tandem mass spectrum is compared with that of the digested peptides of protein database within the mass tolerance that is assigned by users according to the mass spectrometer accuracy. In this study, we used reversed sequence database method to analyze the molecular weight distribution of precursor ions of the tandem mass spectra obtained by the FT LTQ mass spectrometer for human plasma sample. By reinterpreting the precursor ion mass distribution, we could compute the experimental accuracy and we suggested a method to improve the protein identification performance.

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Fermentation and Purification of LacZ-Fused Single Chain Insulin Precursor for($B^{30}$-Homoserine) Human Insulin

  • SeungYup Lee;Jeo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.1 no.1
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    • pp.9-12
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    • 1996
  • In order to produce the single chain precursor of a novel human insulin analogue, (B30-Homoserine) insulin, the fermentative behaviors of Escherichia coli JM103 were studied, which harbors pKBA plasmid carrying a hybrid gene in which the gene for a single chain precursor was fused with lacZ gene under tac promoter. The maximal induction of gene expression was achieved when more than 0.05 mM of isopropyl-$\beta$-D-thiogalactopyranoside(IPTG) was supplemented to fermentation medium after 4 h cultivation of E. coli, and followed by longer than 2-h fermentation. The hybrid protein of the single chain insulin precursor was isolated from cytoplasmic inclusion bodies by dissolving in 8M urea solution, and purified through DEAE-Sephacel and Sephadex G-200 column chromatographies with a recovery of 35%. The finally purified hybrid protein showed a single band on sodium dodecyl sulfate-polyacrylamide gel.

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The Role of Proprotein Convertases in Upper Airway Remodeling

  • Lee, Sang-Nam;Yoon, Joo-Heon
    • Molecules and Cells
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    • v.45 no.6
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    • pp.353-361
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    • 2022
  • Chronic rhinosinusitis (CRS) is a multifactorial, heterogeneous disease characterized by persistent inflammation of the sinonasal mucosa and tissue remodeling, which can include basal/progenitor cell hyperplasia, goblet cell hyperplasia, squamous cell metaplasia, loss or dysfunction of ciliated cells, and increased matrix deposition. Repeated injuries can stimulate airway epithelial cells to produce inflammatory mediators that activate epithelial cells, immune cells, or the epithelial-mesenchymal trophic unit. This persistent inflammation can consequently induce aberrant tissue remodeling. However, the molecular mechanisms driving disease within the different molecular CRS subtypes remain inadequately characterized. Numerous secreted and cell surface proteins relevant to airway inflammation and remodeling are initially synthesized as inactive precursor proteins, including growth/differentiation factors and their associated receptors, enzymes, adhesion molecules, neuropeptides, and peptide hormones. Therefore, these precursor proteins require post-translational cleavage by proprotein convertases (PCs) to become fully functional. In this review, we summarize the roles of PCs in CRS-associated tissue remodeling and discuss the therapeutic potential of targeting PCs for CRS treatment.

Intracellular Posttranslational Modification of Aspartyl Proteinase of Candida albicans and the Role of the Glycan Region of the Enzyme

  • 나병국;송철용
    • Korean Journal of Microbiology
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    • v.38 no.4
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    • pp.218-218
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    • 2002
  • Using two drugs, tunicamycin and brefeldin A, which affect protein processing, we investigated the intracellular processing mechanism of secreted aspartyl proteinase 1 (SAPl) of Candide albicans. Three intracellular forms of SAPI were detected by immunoblotting using menoclonal antibody (MAb) CAPl. Their molecular weights were approximately 40, 41 and 45 kDa, respectively. The 41 kDa protein is a glycoprotein and may be the same as the extracellular form judging by its molecular mass. The 40 kDa protein was the unglycosylated form and its molecular mass coincided with deglycosylated SAPl and the 45 kDa protein was also the unglycosylated form. Neither the 40 and 45 kDa proteins were detected in the culture supernatant of C. albicans. These suggested that the 40 and 45 kDa proteins might be intracellular precursor forms of SAPI. These results show that SAPI is translated as a 45 kDa precusor form in the endoplasmic reticulum and the 45 kDa precursor farm undergoes proteolytic cleavage after translocation into the Golgi apparatus, generating the 40 kDa precursor form. This 40 kDa precursor is converted into a 41 kDa mature form through glycosylation in the Golgi apparatus. The mature form of the 41 kDa protein is sorted into secretary vesicles and finally released into the extracellular space through membrane fusion. When the glycan region of SAPl was digested with N-glycosidase F, both stability and activity of the enzyme decreased. These results indicate that the glycan attached to the enzyme may, at least in parti be related to enzyme stability and activity.

Intracellular Posttranslational Modification of Aspartyl Proteinase of Candida albicans and the Role of the Glycan Region of the Enzyme

  • Na, Byung-Kuk;Song, Chul-Yong
    • Journal of Microbiology
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    • v.38 no.4
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    • pp.218-223
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    • 2000
  • Using two drugs, tunicamycin and brefeldin A, which affect protein processing, we investigated the intracellular processing mechanism of secreted aspartyl proteinase 1 (SAPl) of Candide albicans. Three intracellular forms of SAPI were detected by immunoblotting using menoclonal antibody (MAb) CAPl. Their molecular weights were approximately 40, 41 and 45 kDa, respectively. The 41 kDa protein is a glycoprotein and may be the same as the extracellular form judging by its molecular mass. The 40 kDa protein was the unglycosylated form and its molecular mass coincided with deglycosylated SAPl and the 45 kDa protein was also the unglycosylated form. Neither the 40 and 45 kDa proteins were detected in the culture supernatant of C. albicans. These suggested that the 40 and 45 kDa proteins might be intracellular precursor forms of SAPI. These results show that SAPI is translated as a 45 kDa precusor form in the endoplasmic reticulum and the 45 kDa precursor farm undergoes proteolytic cleavage after translocation into the Golgi apparatus, generating the 40 kDa precursor form. This 40 kDa precursor is converted into a 41 kDa mature form through glycosylation in the Golgi apparatus. The mature form of the 41 kDa protein is sorted into secretary vesicles and finally released into the extracellular space through membrane fusion. When the glycan region of SAPl was digested with N-glycosidase F, both stability and activity of the enzyme decreased. These results indicate that the glycan attached to the enzyme may, at least in parti be related to enzyme stability and activity.

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Pyrolytic Carbon Membranes for Air Separations (공기 분리용 열분해 탄소막)

  • Singh, Anshu;Koros, W.J.
    • Membrane Journal
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    • v.7 no.1
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    • pp.15-21
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    • 1997
  • Carbon molecular sieve (CMS) membranes were synthesized by the pyrolysis of polymeric precursors. The CMS materials had oxygen-nitrogen selectivities much higher than those observed for the polymeric precursors. Typically molecular sieving materials have diffusion selectivities much higher than polymeric materials. This has been identified as a result of higher entropic selectivity of the molecular sieving materials. A study of the development of molecular sieving properties as the polymeric precursor is pyrolyzed into a CMS material will offer us an insight into polymeric molecular structures needed for enhanced entropic selectivity membrane materials.

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Controlling the initial conditions of precursor powders and its effects on the phase evolution and $J_c$ properties of Bi-2223/Ag tapes

  • Jiang, C.H;Yoo, J.M;Kim, H.D;Kang, S.C;Chung, H.S;Wang, Y.Z;Ko, J.W;Qiao, G.W
    • Progress in Superconductivity
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    • v.3 no.1
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    • pp.91-94
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    • 2001
  • By varying fabricating process, precursor powders with different initial conditions were prepared. Subsequently, Bi-2223/Ag tapes were made through these powders. The effects of precursor powders on the phase evolution and Jr properties of Bi-2223/Ag tapes were studied along with several thermomechanical cycles. Our results showed that the initial conditions of precursor powders could strongly influence the phase formation rate and $J_{c}$ value in final tapes. The factors of precursor powders that influence the phase formation and $J_{c}$ of Bi-2223/Ag tapes must be studied and optimized in combination.ion.

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Synthesis of α-oximinoketones, Precursor of CO2 Reduction Macrocyclic Coenzyme F430 Model Complexes

  • Kim, Gilhoon;Won, Hoshik
    • Journal of the Korean Magnetic Resonance Society
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    • v.21 no.4
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    • pp.139-144
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    • 2017
  • Ni(II) containing coenzyme F430 catalyzes the reduction of $CO_2$ in methanogen. Macrocyclic Ni(II) complexes with N,O shiff bases have been received a great attention since metal ions play an important role in the catalysis of reduction. The reducing power of metal complexes are supposed to be dependent on oxidoreduction state of metal ion and structural properties of macrocyclic ring moiety that can enhance electrochemical properties in catalytic process. Six different ${\alpha}$-oximinoketone compounds, precursor of macrocyclic ligands used in $CO_2$ reduction coenzyme F430 model complexes, were synthesized with yields over 90% and characterized by NMR. The molecular geometries of ${\alpha}$-oximinoketone analogues were fully optimized at Beck's-three-parameter hybrid (B3LYP) method in density functional theory (DFT) method with $6-31+G^*$ basis set using the ab initio program. In order to understand molecular planarity and substitutional effects that may enhance reducing power of metal ion are studied by computing the structure-dependent $^{13}C$-NMR chemical shift and comparing with experimental results.