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Influence of protein (human galectin-3) design on aspects of lectin activity
The concept of biomedical significance of the functional pairing between tissue lectins and their glycoconjugate counterreceptors has reached the mainstream of research on the flow of biological information. A major challenge now is to identify the principles of structure–activity relationships that...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Berlin Heidelberg
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7429544/ https://www.ncbi.nlm.nih.gov/pubmed/32335744 http://dx.doi.org/10.1007/s00418-020-01859-9 |
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author | García Caballero, Gabriel Beckwith, Donella Shilova, Nadezhda V. Gabba, Adele Kutzner, Tanja J. Ludwig, Anna-Kristin Manning, Joachim C. Kaltner, Herbert Sinowatz, Fred Cudic, Mare Bovin, Nicolai V. Murphy, Paul V. Gabius, Hans-Joachim |
author_facet | García Caballero, Gabriel Beckwith, Donella Shilova, Nadezhda V. Gabba, Adele Kutzner, Tanja J. Ludwig, Anna-Kristin Manning, Joachim C. Kaltner, Herbert Sinowatz, Fred Cudic, Mare Bovin, Nicolai V. Murphy, Paul V. Gabius, Hans-Joachim |
author_sort | García Caballero, Gabriel |
collection | PubMed |
description | The concept of biomedical significance of the functional pairing between tissue lectins and their glycoconjugate counterreceptors has reached the mainstream of research on the flow of biological information. A major challenge now is to identify the principles of structure–activity relationships that underlie specificity of recognition and the ensuing post-binding processes. Toward this end, we focus on a distinct feature on the side of the lectin, i.e. its architecture to present the carbohydrate recognition domain (CRD). Working with a multifunctional human lectin, i.e. galectin-3, as model, its CRD is used in protein engineering to build variants with different modular assembly. Hereby, it becomes possible to compare activity features of the natural design, i.e. CRD attached to an N-terminal tail, with those of homo- and heterodimers and the tail-free protein. Thermodynamics of binding disaccharides proved full activity of all proteins at very similar affinity. The following glycan array testing revealed maintained preferential contact formation with N-acetyllactosamine oligomers and histo-blood group ABH epitopes irrespective of variant design. The study of carbohydrate-inhibitable binding of the test panel disclosed up to qualitative cell-type-dependent differences in sections of fixed murine epididymis and especially jejunum. By probing topological aspects of binding, the susceptibility to inhibition by a tetravalent glycocluster was markedly different for the wild-type vs the homodimeric variant proteins. The results teach the salient lesson that protein design matters: the type of CRD presentation can have a profound bearing on whether basically suited oligosaccharides, which for example tested positively in an array, will become binding partners in situ. When lectin-glycoconjugate aggregates (lattices) are formed, their structural organization will depend on this parameter. Further testing (ga)lectin variants will thus be instrumental (i) to define the full range of impact of altering protein assembly and (ii) to explain why certain types of design have been favored during the course of evolution, besides opening biomedical perspectives for potential applications of the novel galectin forms. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00418-020-01859-9) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-7429544 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-74295442020-08-19 Influence of protein (human galectin-3) design on aspects of lectin activity García Caballero, Gabriel Beckwith, Donella Shilova, Nadezhda V. Gabba, Adele Kutzner, Tanja J. Ludwig, Anna-Kristin Manning, Joachim C. Kaltner, Herbert Sinowatz, Fred Cudic, Mare Bovin, Nicolai V. Murphy, Paul V. Gabius, Hans-Joachim Histochem Cell Biol Original Paper The concept of biomedical significance of the functional pairing between tissue lectins and their glycoconjugate counterreceptors has reached the mainstream of research on the flow of biological information. A major challenge now is to identify the principles of structure–activity relationships that underlie specificity of recognition and the ensuing post-binding processes. Toward this end, we focus on a distinct feature on the side of the lectin, i.e. its architecture to present the carbohydrate recognition domain (CRD). Working with a multifunctional human lectin, i.e. galectin-3, as model, its CRD is used in protein engineering to build variants with different modular assembly. Hereby, it becomes possible to compare activity features of the natural design, i.e. CRD attached to an N-terminal tail, with those of homo- and heterodimers and the tail-free protein. Thermodynamics of binding disaccharides proved full activity of all proteins at very similar affinity. The following glycan array testing revealed maintained preferential contact formation with N-acetyllactosamine oligomers and histo-blood group ABH epitopes irrespective of variant design. The study of carbohydrate-inhibitable binding of the test panel disclosed up to qualitative cell-type-dependent differences in sections of fixed murine epididymis and especially jejunum. By probing topological aspects of binding, the susceptibility to inhibition by a tetravalent glycocluster was markedly different for the wild-type vs the homodimeric variant proteins. The results teach the salient lesson that protein design matters: the type of CRD presentation can have a profound bearing on whether basically suited oligosaccharides, which for example tested positively in an array, will become binding partners in situ. When lectin-glycoconjugate aggregates (lattices) are formed, their structural organization will depend on this parameter. Further testing (ga)lectin variants will thus be instrumental (i) to define the full range of impact of altering protein assembly and (ii) to explain why certain types of design have been favored during the course of evolution, besides opening biomedical perspectives for potential applications of the novel galectin forms. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00418-020-01859-9) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2020-04-25 2020 /pmc/articles/PMC7429544/ /pubmed/32335744 http://dx.doi.org/10.1007/s00418-020-01859-9 Text en © The Author(s) 2020 Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Original Paper García Caballero, Gabriel Beckwith, Donella Shilova, Nadezhda V. Gabba, Adele Kutzner, Tanja J. Ludwig, Anna-Kristin Manning, Joachim C. Kaltner, Herbert Sinowatz, Fred Cudic, Mare Bovin, Nicolai V. Murphy, Paul V. Gabius, Hans-Joachim Influence of protein (human galectin-3) design on aspects of lectin activity |
title | Influence of protein (human galectin-3) design on aspects of lectin activity |
title_full | Influence of protein (human galectin-3) design on aspects of lectin activity |
title_fullStr | Influence of protein (human galectin-3) design on aspects of lectin activity |
title_full_unstemmed | Influence of protein (human galectin-3) design on aspects of lectin activity |
title_short | Influence of protein (human galectin-3) design on aspects of lectin activity |
title_sort | influence of protein (human galectin-3) design on aspects of lectin activity |
topic | Original Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7429544/ https://www.ncbi.nlm.nih.gov/pubmed/32335744 http://dx.doi.org/10.1007/s00418-020-01859-9 |
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