Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/110437
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Type: Journal article
Title: Structure aided design of a Neu5Gc specific lectin
Author: Day, C.
Paton, A.
Higgins, M.
Shewell, L.
Jen, F.
Schulz, B.
Herdman, B.
Paton, J.
Jennings, M.
Citation: Scientific Reports, 2017; 7(1):1495-1-1495-8
Publisher: Nature Publishing Group
Issue Date: 2017
ISSN: 2045-2322
2045-2322
Statement of
Responsibility: 
Christopher J. Day, Adrienne W. Paton, Melanie A. Higgins, Lucy K. Shewell, Freda E.-C. Jen, Benjamin L. Schulz, Brock P. Herdman, James C. Paton and Michael P. Jennings
Abstract: Subtilase cytotoxin (SubAB) of Escherichia coli is an AB5 class bacterial toxin. The pentameric B subunit (SubB) binds the cellular carbohydrate receptor, α2-3-linked N-glycolylneuraminic acid (Neu5Gc). Neu5Gc is not expressed on normal human cells, but is expressed by cancer cells. Elevated Neu5Gc has been observed in breast, ovarian, prostate, colon and lung cancer. The presence of Neu5Gc is prognostically important, and correlates with invasiveness, metastasis and tumour grade. Neu5Gc binding by SubB suggests that it may have utility as a diagnostic tool for the detection Neu5Gc tumor antigens. Native SubB has 20-fold less binding to N-acetlylneuraminic acid (Neu5Ac); over 30-fold less if the Neu5Gc linkage was changed from α2-3 to α2-6. Using molecular modeling approaches, site directed mutations were made to reduce the α2-3»α2-6-linkage preference, while maintaining or enhancing the selectivity of SubB for Neu5Gc over Neu5Ac. Surface plasmon resonance and glycan array analysis showed that the SubBΔS106/ΔT107 mutant displayed improved specificity towards Neu5Gc and bound to α2-6-linked Neu5Gc. SubBΔS106/ΔT107 could discriminate NeuGc- over Neu5Ac-glycoconjugates in ELISA. These data suggest that improved SubB mutants offer a new tool for the testing of biological samples, particularly serum and other fluids from individuals with cancer or suspected of having cancer.
Keywords: Animals; Cattle; Humans; Neuraminic Acids; Subtilisins; Escherichia coli Proteins; Lectins; Enzyme-Linked Immunosorbent Assay; Surface Plasmon Resonance; Protein Engineering; Binding Sites; Mutation; Models, Molecular; Mutant Proteins
Rights: © The Author(s) 2017 This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
RMID: 0030069322
DOI: 10.1038/s41598-017-01522-9
Grant ID: http://purl.org/au-research/grants/nhmrc/1071659
http://purl.org/au-research/grants/nhmrc/1087975
http://purl.org/au-research/grants/arc/DP160102766
Appears in Collections:Molecular and Biomedical Science publications

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