• Title/Summary/Keyword: Bio-fusion

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A Fusion Tag to Fold on: The S-Layer Protein SgsE Confers Improved Folding Kinetics to Translationally Fused Enhanced Green Fluorescent Protein

  • Ristl, Robin;Kainz, Birgit;Stadlmayr, Gerhard;Schuster, Heinrich;Pum, Dietmar;Messner, Paul;Obinger, Christian;Schaffer, Christina
    • Journal of Microbiology and Biotechnology
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    • v.22 no.9
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    • pp.1271-1278
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    • 2012

  • Genetic fusion of two proteins frequently induces beneficial effects to the proteins, such as increased solubility, besides the combination of two protein functions. Here, we study the effects of the bacterial surface layer protein SgsE from Geobacillus stearothermophilus NRS 2004/3a on the folding of a C-terminally fused enhanced green fluorescent protein (EGFP) moiety. Although GFPs are generally unable to adopt a functional confirmation in the bacterial periplasm of Escherichia coli cells, we observed periplasmic fluorescence from a chimera of a 150-amino-acid N-terminal truncation of SgsE and EGFP. Based on this finding, unfolding and refolding kinetics of different S-layer-EGFP chimeras, a maltose binding protein-EGFP chimera, and sole EGFP were monitored using green fluorescence as indicator for the folded protein state. Calculated apparent rate constants for unfolding and refolding indicated different folding pathways for EGFP depending on the fusion partner used, and a clearly stabilizing effect was observed for the SgsE_C fusion moiety. Thermal stability, as determined by differential scanning calorimetry, and unfolding equilibria were found to be independent of the fused partner. We conclude that the stabilizing effect SgsE_C exerts on EGFP is due to a reduction of degrees of freedom for folding of EGFP in the fused state.

  • https://doi.org/10.4014/jmb.1202.02023 인용 PDF KSCI

Bio-Inspired Object Recognition Using Parameterized Metric Learning

  • Li, Xiong;Wang, Bin;Liu, Yuncai
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • v.7 no.4
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    • pp.819-833
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    • 2013

  • Computing global features based on local features using a bio-inspired framework has shown promising performance. However, for some tough applications with large intra-class variances, a single local feature is inadequate to represent all the attributes of the images. To integrate the complementary abilities of multiple local features, in this paper we have extended the efficacy of the bio-inspired framework, HMAX, to adapt heterogeneous features for global feature extraction. Given multiple global features, we propose an approach, designated as parameterized metric learning, for high dimensional feature fusion. The fusion parameters are solved by maximizing the canonical correlation with respect to the parameters. Experimental results show that our method achieves significant improvements over the benchmark bio-inspired framework, HMAX, and other related methods on the Caltech dataset, under varying numbers of training samples and feature elements.

  • https://doi.org/10.3837/tiis.2013.04.012 인용 PDF KSCI

Effect of Fc Fusion on Folding and Immunogenicity of Middle East Respiratory Syndrome Coronavirus Spike Protein

  • Chun, Jungmin;Cho, Yeondong;Park, Ki Hoon;Choi, Hanul;Cho, Hansam;Lee, Hee-Jung;Jang, Hyun;Kim, Kyung Hyun;Oh, Yu-Kyoung;Kim, Young Bong
    • Journal of Microbiology and Biotechnology
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    • v.29 no.5
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    • pp.813-819
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    • 2019

  • Middle East respiratory syndrome coronavirus (MERS-CoV) induces severe respiratory impairment with a reported mortality rate of ~36% in humans. The absence of clinically available MERS-CoV vaccines and treatments to date has resulted in uncontrolled incidence and propagation of the virus. In vaccine design, fusion with the IgG Fc domain is reported to increase the immunogenicity of various vaccine antigens. However, limited reports have documented the potential negative effects of Fc fusion on vaccine antigens. To determine whether Fc fusion affects the immunogenicity of MERS-CoV antigen, we constructed a Fcassociated MERS-CoV spike protein (eS770-Fc, 110 kDa), whereby human IgG4 Fc domain was fused to MERS-CoV spike protein (eS770) via a Gly/Pro linker using baculovirus as the expression system. For comparative analyses, two eS770 proteins lacking the IgG4 Fc domain were generated using the IdeS protease ($eS770-{\Delta}Fc$) or His tag attachment (eS770-His) and the immunogenicity of the above constructs were examined following intramuscular immunization in mice. Contrary to expectations, non-Fc spike proteins ($eS770-{\Delta}Fc$, eS770-His; 90 kDa) showed higher immunogenicity than the Fc fusion protein (eS770-Fc). Moreover, unlike non-Fc spike proteins, eS770-Fc immunization did not elicit neutralizing antibodies against MERS-CoV. The lower immunogenicity of Fc-fused eS770 was related to alterations in the structural conformation of the spike protein. Taken together, our results indicate that IgG Fc fusion reduces the immunogenicity of eS770 by interfering with the proper folding structure.

  • https://doi.org/10.4014/jmb.1903.03043 인용 PDF KSCI