Nature-inspired engineering of an F-type lectin for increased binding strength.
DIR@IMTECH: CSIR-Institute of Microbial Technology
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Title |
Nature-inspired engineering of an F-type lectin for increased binding strength.
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Creator |
Mahajan, Sonal
Ramya, T N C |
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Subject |
QR Microbiology
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Description |
Individual lectin-carbohydrate interactions are usually of low affinity. However, high avidity is frequently attained by the multivalent presentation of glycans on biological surfaces coupled with the occurrence of high order lectin oligomers or tandem repeats of lectin domains in the polypeptide. F-type lectins are l-fucose binding lectins with a typical sequence motif, HX(26)RXDX(4)R/K, whose residues participate in l-fucose binding. We previously reported the presence of a few eukaryotic F-type lectin domains with partial sequence duplication that results in the presence of two l-fucose-binding sequence motifs. We hypothesized that such partial sequence duplication would result in greater avidity of lectin-ligand interactions. Inspired by this example from Nature, we attempted to engineer a bacterial F-type lectin domain from Streptosporangium roseum to attain avid binding by mimicking partial duplication. The engineered lectin demonstrated 12-fold greater binding strength than the wild-type lectin to multivalent fucosylated glycoconjugates. However, the affinity to the monosaccharide l-fucose in solution was similar and partial sequence duplication did not result in an additional functional l-fucose binding site. We also cloned, expressed and purified a Branchiostoma floridae F-type lectin domain with naturally occurring partial sequence duplication and confirmed that the duplicated region with the F-type lectin sequence motif did not participate in l-fucose binding. We found that the greater binding strength of the engineered lectin from S. roseum was instead due to increased oligomerization. We believe that this Nature-inspired strategy might be useful for engineering lectins to improve binding strength in various applications.
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Publisher |
Oxford University Press
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Date |
2018-12-01
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Type |
Article
PeerReviewed |
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Relation |
https://academic.oup.com/glycob/article/28/12/933/5094506
http://crdd.osdd.net/open/2148/ |
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Identifier |
Mahajan, Sonal and Ramya, T N C (2018) Nature-inspired engineering of an F-type lectin for increased binding strength. Glycobiology, 28 (12). pp. 933-948. ISSN 1460-2423
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