Abstract
Galectins,β-galactoside binding proteins, function in several physiological and pathological processes. The further evaluation of these processes as well as possible applications of galectins in diagnosis and therapy has raised high scientific interest. Therefore, easy and reliable test systems are necessary. Here we present the simple and cost-efficient production of recombinant human galectins as fusion proteins with SNAP-tag and fluorescent proteins. These constructs show binding specificities and oligomerisation properties generally comparable to recombinant galectins. Their direct fluorescence signal was utilised by ELISA-type assay and flow cytometry analysis with human and ovine mesenchymal stem cells (MSC). Flow cytometry demonstrated glycan mediated binding of His6-SNAP-YFP-Gal- 3 to both MSC types, which was specifically inhibited by lactose. Moreover, directed immobilisation by SNAP-tag technology onto benzylguanine- activated sepharose was utilised to prepare galectin affinity columns for glycoprotein analysis and purification. The SNAPtag directed coupling yielded up to three-fold higher binding capacities for the glycoprotein standard asialofetuin compared to nondirected coupled galectin suggesting improved functionality following directed coupling.
Keywords: Galectin-1, galectin-3, SNAP-tag, fusion protein, fluorescent protein, mesenchymal stem cells.
Current Pharmaceutical Design
Title:Fluorescent SNAP-Tag Galectin Fusion Proteins as Novel Tools in Glycobiology
Volume: 19 Issue: 30
Author(s): Christiane E. Kupper, Sophia Böcker, Hulong Liu, Carina Adamzyk, Julia van de Kamp, Tobias Recker, Bernd Lethaus, Willi Jahnen-Dechent, Sabine Neuss, Gerhard Muller-Newen and Lothar Elling
Affiliation:
Keywords: Galectin-1, galectin-3, SNAP-tag, fusion protein, fluorescent protein, mesenchymal stem cells.
Abstract: Galectins,β-galactoside binding proteins, function in several physiological and pathological processes. The further evaluation of these processes as well as possible applications of galectins in diagnosis and therapy has raised high scientific interest. Therefore, easy and reliable test systems are necessary. Here we present the simple and cost-efficient production of recombinant human galectins as fusion proteins with SNAP-tag and fluorescent proteins. These constructs show binding specificities and oligomerisation properties generally comparable to recombinant galectins. Their direct fluorescence signal was utilised by ELISA-type assay and flow cytometry analysis with human and ovine mesenchymal stem cells (MSC). Flow cytometry demonstrated glycan mediated binding of His6-SNAP-YFP-Gal- 3 to both MSC types, which was specifically inhibited by lactose. Moreover, directed immobilisation by SNAP-tag technology onto benzylguanine- activated sepharose was utilised to prepare galectin affinity columns for glycoprotein analysis and purification. The SNAPtag directed coupling yielded up to three-fold higher binding capacities for the glycoprotein standard asialofetuin compared to nondirected coupled galectin suggesting improved functionality following directed coupling.
Export Options
About this article
Cite this article as:
Kupper E. Christiane, Böcker Sophia, Liu Hulong, Adamzyk Carina, Kamp van de Julia, Recker Tobias, Lethaus Bernd, Jahnen-Dechent Willi, Neuss Sabine, Muller-Newen Gerhard and Elling Lothar, Fluorescent SNAP-Tag Galectin Fusion Proteins as Novel Tools in Glycobiology, Current Pharmaceutical Design 2013; 19 (30) . https://dx.doi.org/10.2174/1381612811319300017
DOI https://dx.doi.org/10.2174/1381612811319300017 |
Print ISSN 1381-6128 |
Publisher Name Bentham Science Publisher |
Online ISSN 1873-4286 |
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
- Announcements
Related Articles
-
Gene Therapy (Part II)
Current Gene Therapy A Review on Novel Breast Cancer Therapies: Photodynamic Therapy and Plant Derived Agent Induced Cell Death Mechanisms
Anti-Cancer Agents in Medicinal Chemistry Epigenetic Regulation of ABCB1 Transporter Expression and Function
Current Pharmacogenomics and Personalized Medicine Reduction of Breast Cancer Relapses with Perioperative Non-Steroidal Anti-Inflammatory Drugs: New Findings and a Review
Current Medicinal Chemistry Tumor Necrosis Factor-α-Mediated Pulmonary Endothelial Barrier Dysfunction
Current Respiratory Medicine Reviews Using NMR to Develop New Allosteric and Allo-Network Drugs
Current Drug Discovery Technologies The Scatter Factor Signaling Pathways as Therapeutic Associated Target in Cancer Treatment
Current Medicinal Chemistry Heparin, Heparan Sulfate and Heparanase in Cancer: Remedy for Metastasis?
Anti-Cancer Agents in Medicinal Chemistry Small Molecules as Anti-TNF Drugs
Current Medicinal Chemistry Developments in the Application of 1,2,3-Triazoles in Cancer Treatment
Recent Patents on Anti-Cancer Drug Discovery Galectin-9 in Cancer Therapy
Recent Patents on Endocrine, Metabolic & Immune Drug Discovery (Discontinued) Role of Cytomegalovirus in Salivary Gland Tumors: An Unfolded Mystery
Recent Patents on Biomarkers Disruption of Metabolic Pathways - Perspectives for the Treatment of Cancer
Current Cancer Drug Targets Immunotherapy Approaches in Cancer Treatment
Current Pharmaceutical Biotechnology Vascular Mimicry: Concepts and Implications for Anti-Angiogenic Therapy
Current Angiogenesis (Discontinued) MicroRNA as Regulators of Cancer Stem Cells and Chemoresistance in Colorectal Cancer
Current Cancer Drug Targets Macromolecular and Nanotechnological Modification of Camptothecin and its Analogs to Improve the Efficacy
Current Drug Discovery Technologies Tumour-Specific Uptake of Anti-Cancer Drugs: The Future is Here
Current Drug Metabolism Perspectives in Nanomedicine-Based Research Towards Cancer Therapies
Current Nanoscience Secretory Clusterin as a Novel Molecular-targeted Therapy for Inhibiting Hepatocellular Carcinoma Growth
Current Medicinal Chemistry