Generation of bivalent and bispecific kringle single domains by loop grafting as potent agonists against death receptors 4 and 5 = 이가-이중 특이 크링글 도메인 단량체 유도

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dc.contributor.authorC H Lee-
dc.contributor.authorK J Park-
dc.contributor.authorSang Jick Kim-
dc.contributor.authorOh Suk Kwon-
dc.contributor.authorK J Jeong-
dc.contributor.authorA Kim-
dc.contributor.authorY S Kim-
dc.description.abstractBivalent or bispecific binding activity of proteins has been mainly achieved by assembling two or more domains in a single molecule. Here we report bivalent/bispecific single-domain proteins based on the kringle domain (KD), which has a cystine knot structural motif and is highly tolerant of sequence modifications. KD has seven loops protruding from the core fold into two largely opposite directions, dubbed loop cluster regions (LCRs) 1 and 2. Mutational analysis of previously isolated agonistic KD variants against human death receptors (DRs) 4 and 5 revealed that they can simultaneously recognize two target molecules of DR4 and/or DR5 via the two independent binding sites of LCR1 and LCR2. Binding loop mapping of yeast-surface-displayed KD mutants identified high-affinity target binding loops in LCR2, which were then grafted into conformationally compatible loops located on the opposite side of LCR1 within the same or different KD variants to generate bivalent/bispecific KD variants against DR4 and/or DR5 with improved affinity. The loop-grafted bivalent/bispecific KD variants showed enhanced cell-death-inducing activity of tumor cells compared with their monovalent/monospecific and bivalent/ monospecific counterparts, demonstrating an advantage of bispecific targeting to both DR4 and DR5 over the targeting of only one of the two pro-apoptotic receptors. Our results suggest that the KD with the two independent binding surfaces for target recognition is an appropriate scaffold for the development of bivalency and/or bispecificity by loop grafting on the single domain, which offers a distinct advantage over other protein scaffolds with a single binding surface.-
dc.titleGeneration of bivalent and bispecific kringle single domains by loop grafting as potent agonists against death receptors 4 and 5 = 이가-이중 특이 크링글 도메인 단량체 유도-
dc.title.alternativeGeneration of bivalent and bispecific kringle single domains by loop grafting as potent agonists against death receptors 4 and 5-
dc.citation.titleJournal of Molecular Biology-
dc.contributor.affiliatedAuthorSang Jick Kim-
dc.contributor.affiliatedAuthorOh Suk Kwon-
dc.identifier.bibliographicCitationJournal of Molecular Biology, vol. 411, no. 1, pp. 201-219-
dc.subject.keywordcell death-
dc.subject.keywordnonantibody protein scaffold-
dc.subject.keywordprotein engineering-
dc.subject.keywordTRAIL receptor-
dc.subject.keywordyeast surface display-
dc.subject.localcell death-
dc.subject.localCell death-
dc.subject.localnonantibody protein scaffold-
dc.subject.localProtein engineering-
dc.subject.localprotein engineering-
dc.subject.localProtein Engineering-
dc.subject.localTRAIL receptor-
dc.subject.localyeast surface display-
dc.subject.localYeast surface display-
Appears in Collections:
Synthetic Biology and Bioengineering Research Institute > Synthetic Biology Research Center > 1. Journal Articles
Division of Bio Technology Innovation > SME Support Center > 1. Journal Articles
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