High-Affinity N ‑(2-Hydroxypropyl)methacrylamide Copolymers with Tailored N ‑Acetyllactosamine Presentation Discriminate between Galectins

Biomacromolecules, vol. 21, pp. 641–652

Abstract

Abstract N-Acetyllactosamine (LacNAc; Galβ4GlcNAc) is a typical disaccharide ligand of galectins. The most abundant members of these human lectins, galectin-1 (Gal-1) and galectin-3 (Gal-3), participate in a number of pathologies including cancerogenesis and metastatic formation. In this study, we synthesized a series of fifteen N-(2-hydroxypropyl)­methacrylamide (HPMA)-based glycopolymers with varying LacNAc amounts and presentations and evaluated the impact of their architecture on the binding affinity to Gal-1 and Gal-3. The controlled radical reversible addition–fragmentation chain transfer copolymerization technique afforded linear polymer precursors with comparable molecular weight (Mn ≈ 22,000 g mol–1) and narrow dispersity (D̵ ≈ 1.1). The precursors were conjugated with the functionalized LacNAc disaccharide (4–22 mol % content in glycopolymer) prepared by enzymatic synthesis under catalysis by β-galactosidase from Bacillus circulans. The structure–affinity relationship study based on the enzyme-linked immunosorbent assay revealed that the type of LacNAc presentation, individual or clustered on bi- or trivalent linkers, brings a clear discrimination (almost 300-fold) between Gal-1 and Gal-3, reaching avidity to Gal-1 in the nanomolar range. Whereas Gal-1 strongly preferred a dense presentation of individually distributed LacNAc epitopes, Gal-3 preferred a clustered LacNAc presentation. Such a strong galectin preference based just on the structure of a multivalent glycopolymer type is exceptional. The prepared nontoxic, nonimmunogenic, and biocompatible glycopolymers are prospective for therapeutic applications requiring selectivity for one particular galectin.

Authors 10

1. Marina Rodrigues Tavares

   Czech Academy of Sciences · Czech Academy of Sciences, Institute of Macromolecular Chemistry

   Affiliation as printed

   Czech Academy of Sciences , , Heyrovského náměstí 2 , ,

   Institute of Macromolecular Chemistry, Czech Academy of Sciences, Heyrovského náměstí 2, CZ-162 06 Prague 6, Czech Republic

2. Markéta Bláhová

   Czech Academy of Sciences · Czech Academy of Sciences, Institute of Macromolecular Chemistry

   Affiliation as printed

   Czech Academy of Sciences , , Heyrovského náměstí 2 , ,

   Institute of Macromolecular Chemistry, Czech Academy of Sciences, Heyrovského náměstí 2, CZ-162 06 Prague 6, Czech Republic

3. Lieselotte Sedláková

   Czech Academy of Sciences · Czech Academy of Sciences, Institute of Microbiology · Czech Technical University in Prague

   Affiliation as printed

   Czech Academy of Sciences , , Vídeňská 1083 , ,

   Czech Technical University in Prague , , Sítná sq. 3105 , ,

   Department of Health Care Disciplines and Population Protection, Faculty of Biomedical Engineering, Czech Technical University in Prague, Sítná sq. 3105, CZ-272 01 Kladno, Czech Republic

   Institute of Microbiology, Czech Academy of Sciences, Vídeňská 1083, CZ-142 20 Prague 4, Czech Republic

4. Lothar Elling Aachen

   RWTH Aachen University

   Affiliation as printed

   RWTH Aachen , , Pauwelstr. 20 , ,

   Institute of Biotechnology and Helmholtz Institute for Biomedical Engineering, RWTH Aachen, Pauwelstr. 20, D-52079 Aachen, Germany

5. Helena Pelantová

   Czech Academy of Sciences · Czech Academy of Sciences, Institute of Microbiology

   Affiliation as printed

   Czech Academy of Sciences , , Vídeňská 1083 , ,

   Institute of Microbiology, Czech Academy of Sciences, Vídeňská 1083, CZ-142 20 Prague 4, Czech Republic

6. Rafał Konefał

   Czech Academy of Sciences · Czech Academy of Sciences, Institute of Macromolecular Chemistry

   Affiliation as printed

   Czech Academy of Sciences , , Heyrovského náměstí 2 , ,

   Institute of Macromolecular Chemistry, Czech Academy of Sciences, Heyrovského náměstí 2, CZ-162 06 Prague 6, Czech Republic

7. Tomáš Etrych

   Czech Academy of Sciences · Czech Academy of Sciences, Institute of Macromolecular Chemistry

   Affiliation as printed

   Czech Academy of Sciences , , Heyrovského náměstí 2 , ,

   Institute of Macromolecular Chemistry, Czech Academy of Sciences, Heyrovského náměstí 2, CZ-162 06 Prague 6, Czech Republic

8. Vladimı́r Křen

   Czech Academy of Sciences · Czech Academy of Sciences, Institute of Microbiology

   Affiliation as printed

   Czech Academy of Sciences , , Vídeňská 1083 , ,

   Institute of Microbiology, Czech Academy of Sciences, Vídeňská 1083, CZ-142 20 Prague 4, Czech Republic

9. Pavla Bojarová corresponding

   Czech Academy of Sciences · Czech Academy of Sciences, Institute of Microbiology · Czech Technical University in Prague

   Affiliation as printed

   Czech Academy of Sciences , , Vídeňská 1083 , ,

   Czech Technical University in Prague , , Sítná sq. 3105 , ,

   Department of Health Care Disciplines and Population Protection, Faculty of Biomedical Engineering, Czech Technical University in Prague, Sítná sq. 3105, CZ-272 01 Kladno, Czech Republic

   Institute of Microbiology, Czech Academy of Sciences, Vídeňská 1083, CZ-142 20 Prague 4, Czech Republic

10. Petr Chytil corresponding

    Czech Academy of Sciences · Czech Academy of Sciences, Institute of Macromolecular Chemistry

    Affiliation as printed

    Czech Academy of Sciences , , Heyrovského náměstí 2 , ,

    Institute of Macromolecular Chemistry, Czech Academy of Sciences, Heyrovského náměstí 2, CZ-162 06 Prague 6, Czech Republic