title: Effects of the Fc-III tag on activity and stability of green fluorescent protein and human muscle creatine kinase.
abstract: The Fc-III tag is a newly developed fusion tag that can be applied to protein purification and detection. In the present work, we use the Fc-III-tagged green fluorescent protein (GFP) and human muscle creatine kinase (CK) as model systems to investigate effects of the Fc-III tag on activities and stabilities of the expressed multicysteine-containing proteins. Our results show the Fc-III tag has no adverse effects on the fluorescence of GFP and reduces the occurrence of GFP misfolding due to incorrect Cys oxidation compared with the His-tagged protein. The activity and stability of the Fc-III-tagged CK is slightly lower than that of the tag-free CK, but is higher than that of the His-tagged CK as determined by the ratio of the oxidized versus reduced CK. A major portion of His-tagged CK is in its oxidized form, while that of the Fc-III-tagged CK is in its reduced form. A folding model of CK with different tags was proposed, which may provide insights into the effect of the Fc-III tag on the conformations of disulfide-bridged proteins.
authors: MOE Key Laboratory of Bioinformatics, School of Life Sciences, Tsinghua University, Beijing, China.
authors: Shan
authors: S
authors: Feng
authors: Yiyi
authors: Y
authors: Gong
authors: Gulishana
authors: G
authors: Adilijiang
authors: Haiteng
authors: H
authors: Deng
mesh_terms: D052283:Creatine Kinase, MM Form / Q000737:chemistry* / Q000378:metabolism; D004220:Disulfides / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000737:chemistry* / Q000378:metabolism; D010084:Oxidation-Reduction; D058849:Protein Refolding; D055550:Protein Stability; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000378:metabolism; D013696:Temperature
chemical_list: D004220:Disulfides; D007141:Immunoglobulin Fc Fragments; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D052283:Creatine Kinase, MM Form
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