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Protein–carbohydrate interactions
At the heart of biochemistry
Proteins that possess the ability to bind carbohydrates specifically and in the micromolar range. Biologically by Nathan Sharon
reversibly abound in nature, being present in all living organisms, from relevant interaction often requires (The Weizmann Institute of
viruses to humans. Their interactions with their ligands are the basis multivalency of both protein and Science, Rehovot, Israel)
of a myriad of biological processes, both normal and pathological1–3 ligand in order to generate higher
(Table 1). The high selectivity required for these interactions is affinity of binding. Although pro-
provided by a specific stereochemical fit between complementary teins specific for mono-, di- or
molecules, the protein on the one hand and the carbohydrate on the tri-saccharides have small combining
other. This concept has its origins in the lock-and-key hypothesis, sites (Figures 1 and 2), they can read-
introduced by Emil Fischer at the end of the 19th Century to explain ily bind higher oligosaccharides or
the specificity of interactions between enzymes (he studied their derived glycoconjugates. The
glycosidases) and their substrates (carbohydrates), i.e. between combining sites of polysaccharide-
molecules in solution. In time it was extended to describe the specific proteins are not large either,
interactions of cells with soluble molecules and with other cells. and usually do not exceed the size of
approximately six monosaccharide
Complex carbohydrates are found monosaccharides to polysaccharides, units. For example, the binding sites
commonly at the cell surface, where either free (such as starch, cellulose or of lysozyme and of Taka-amylase
they are positioned to interact with hyaluronic acid) or conjugated. The (which hydrolyses amylose) have six Figure 1.
suitable proteinaceous receptors, affinity of such proteins for mono- and seven adjacent subsites respec- N-Acetyllactosamine
primarily lectins, in solution or on saccharides is generally low, in the tively, each complementary to a (Galβ1-4GlcNAc) in the
4,5
the surfaces of other cells . These millimolar range, while for oligosac- monosaccharide; phosphorylase combining site of Erythrina
proteins, originally identified as charides or polysaccharides it may be (which acts on glycogen) has four corallodendron lectin
sugar-specific haemagglutinins, are (a legume seed lectin).
currently known as cell-recognition The essential contact
molecules. They are geared to distin- residues, which include
guish between different oligosaccha- Phe131 (F131) on which
rides, whether as such or as part of the galactose of the ligand
glycoconjugates, most commonly is stacked, are marked. The
glycoproteins and glycolipids. blue sphere denotes the
water molecule that
Combining sites mediates the interaction
between the main chain
Carbohydrate-binding proteins, amide of Leu86 (L86) and
whether enzymes, anti-carbohydrate the O-6 of the galactose.
antibodies, sugar transporters, Broken lines denote
growth factors or lectins, are struc- hydrogen bonds. The cavity
turally diverse, differing markedly in marked with an asterisk is
size and tertiary and quaternary where bulky substituents,
structure. They can bind a wide spec- such as fucose, attached to
trum of ligands, ranging from simple O-2 of the galactose, can
bind (PDB entry 1LTE).
Key words: carbohydrate-binding protein, Courtesy of Miri Eisenstein,
haemagglutinin, ligand recognition, saccharide, Weizmann Institute
X-ray crystallography. of Science
such subsites; the FGF (fibroblast bining sites of carbohydrate-binding detailed atomic picture of the com-
growth factor) combining site fits a proteins was quite limited. It was plexes in the crystal, which appears
particular hexasaccharide sequence of based almost exclusively on speci- to reflect their structure in solution.
heparan sulphate (Figure 3); and the ficity studies of carbohydrate-spe- It was first applied in the 1960s by
combining site of an antibody against cific enzymes and antibodies, David C. Phillips and co-workers
a bacterial polysaccharide possesses complemented by selective chemical to determine the three-dimensional
Figure 2. Combining site of five monosaccharide-specific subsites modification of the proteins. At structure of complexes of hen egg-
human influenza virus (Figure 4), whereas those of antibod- present, the major source of infor- white lysozyme with oligosaccha-
haemagglutinin in complex ies against dextran (a polymer of mation about such sites of carbohy- rides derived from chitin or from
with the trisaccharide α-glucose) can accommodate up to drate-binding proteins is X-ray bacterial cell wall peptidoglycan,
receptor NeuAcα2-3Galβ1- seven monosaccharide units. crystallography of their complexes both substrates of the enzyme. To
4GlcNAc. Hydrogen bonds Early information on the com- with ligands. The method gives a date, the structures of hundreds of
are indicated by broken protein–carbohydrate complexes
lines; amino acid residues have been elucidated, a large number
making interactions via Asn-193 of these of being lectins (see
main-chain carbonyl groups 1934-human/human www.cermav.cnrs.fr/lectines)
receptor complex
are shown as red spheres, (Figure 1). Other sources of combin-
and those interacting via ing-site information include binding
190-Helix
main chain nitrogens are GlcNAc-3 experiments with different sugars
shown as blue spheres. The Glu-190 and their derivatives (a much wider
trisaccharide carbon atoms range than in the past) using state of
are in yellow, its nitrogen Gal-2 the art techniques, site-directed
atoms in blue and oxygen 2 mutagenesis of the proteins and also,
atoms in red. Water 3 to a limited extent, NMR experi-
Lys-222
molecules are indicated by 4 ments and molecular modelling.
Sialic acid
green spheres. Reproduced Such studies have shown that,
227
with permission from like the proteins themselves, the
135
Gamblin, S.J., Haire, L.F., combining sites are diverse, even
Asp-225 Gln-226
Russell, R.J. et al. (2004) 130-Loop when their specificity is the same
220-Loop Thr-136
Science 303, 1838–1842. 137 (although within a given protein
Copyright 2004 AAAS. family, the sites may be similar).