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dc.contributor.author윤형돈ko
dc.contributor.authorGovindan Raghunathan[Govindan Raghunathan]ko
dc.contributor.authorNagasundarapandian Soundrarajan[Nagasundarapandian Soundrarajan]ko
dc.contributor.authorSriram Sokalingam[Sriram Sokalingam]ko
dc.contributor.author이선구[이선구]ko
dc.date.accessioned2015-12-17T02:00:43Z-
dc.date.available2015-12-17T02:00:43Z-
dc.date.created2015-11-13-
dc.date.issued201212-
dc.identifier.citationPLOS ONE, v.7, no.12-
dc.identifier.issn1932-6203-
dc.identifier.urihttp://hdl.handle.net/YU.REPOSITORY/26680-
dc.identifier.urihttp://dx.doi.org/10.1371/journal.pone.0051510-
dc.description.abstractDiversification of protein sequence-structure space is a major concern in protein engineering. Deletion mutagenesis can generate a protein sequence-structure space different from substitution mutagenesis mediated space, but it has not been widely used in protein engineering compared to substitution mutagenesis, because it causes a relatively huge range of structural perturbations of target proteins which often inactivates the proteins. In this study, we demonstrate that, using green fluorescent protein (GFP) as a model system, the drawback of the deletional protein engineering can be overcome by employing the protein structure with high stability. The systematic dissection of N-terminal, C-terminal and internal sequences of GFPs with two different stabilities showed that GFP with high stability (s-GFP), was more tolerant to the elimination of amino acids compared to a GFP with normal stability (n-GFP). The deletion studies of s-GFP enabled us to achieve three interesting variants viz. s-DL4, s-N14, and s-C225, which could not been obtained from n-GFP. The deletion of 191-196 loop sequences led to the variant s-DL4 that was expressed predominantly as insoluble form but mostly active. The s-N14 and s-C225 are the variants without the amino acid residues involving secondary structures around N- and C-terminals of GFP fold respectively, exhibiting comparable biophysical properties of the n-GFP. Structural analysis of the variants through computational modeling study gave a few structural insights that can explain the spectral properties of the variants. Our study suggests that the protein sequence-structure space of deletion mutants can be more efficiently explored by employing the protein structure with higher stability.-
dc.language영어-
dc.publisherPUBLIC LIBRARY SCIENCE-
dc.subjectGREEN-FLUORESCENT PROTEIN-
dc.subjectSECONDARY STRUCTURE ANALYSES-
dc.subjectSULFOLOBUS-SOLFATARICUS-
dc.subjectNATIVE-LIKE-
dc.subjectCRYSTAL-STRUCTURE-
dc.subjectINCLUSION-BODIES-
dc.subjectSEQUENCE SPACE-
dc.subjectSTABILITY-
dc.subjectEVOLUTION-
dc.subjectACYLPHOSPHATASE-
dc.titleDeletional Protein Engineering Based on Stable Fold-
dc.typeArticle-
dc.identifier.wosid000312290800070-
dc.identifier.scopusid2-s2.0-84871156326-
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