Dissecting the cholera toxin-ganglioside GM1 interaction by isothermal titration calorimetry.

The complex of cholera toxin and ganglioside GM1 is one of the highest affinity protein-carbohydrate interactions known. Herein, the GM1 pentasaccharide is dissected into smaller fragments to determine the contribution of each of the key monosaccharide residues to the overall binding affinity. Displacement isothermal titration calorimetry (ITC) has allowed the measurement of all of the key thermodynamic parameters for even the lowest affinity fragment ligands. Analysis of the standard free energy changes using Jencks' concept of intrinsic free energies reveals that the terminal galactose and sialic acid residues contribute 54% and 44% of the intrinsic binding energy, respectively, despite the latter ligand having little appreciable affinity for the toxin. This analysis also provides an estimate of 25.8 kJ mol(-1) for the loss of independent translational and rotational degrees of freedom on complexation and presents evidence for an alternative binding mode for ganglioside GM2. The high affinity and selectivity of the GM1-cholera toxin interaction originates principally from the conformational preorganization of the branched pentasaccharide rather than through the effect of cooperativity, which is also reinvestigated by ITC.

[1]  A. Bernardi,et al.  Mimicking Gangliosides by Design: Mimics of GM1 Headgroup , 2002, Neurochemical Research.

[2]  W Bruce Turnbull,et al.  On the value of c: can low affinity systems be studied by isothermal titration calorimetry? , 2003, Journal of the American Chemical Society.

[3]  S. Homans,et al.  Large-scale millisecond intersubunit dynamics in the B subunit homopentamer of the toxin derived from Escherichia coli O157. , 2003, Journal of the American Chemical Society.

[4]  C. Schengrund.  "Multivalent" saccharides: development of new approaches for inhibiting the effects of glycosphingolipid-binding pathogens. , 2003, Biochemical pharmacology.

[5]  D. Potenza,et al.  Mimics of ganglioside GM1 as cholera toxin ligands: replacement of the GalNAc residue. , 2003, Organic & biomolecular chemistry.

[6]  W. Hol,et al.  Characterization and crystal structure of a high-affinity pentavalent receptor-binding inhibitor for cholera toxin and E. coli heat-labile enterotoxin. , 2002, Journal of the American Chemical Society.

[7]  C. Locht,et al.  Enhanced bacterial virulence through exploitation of host glycosaminoglycans , 2002, Molecular microbiology.

[8]  C. Verlinde,et al.  Anchor-based design of improved cholera toxin and E. coli heat-labile enterotoxin receptor binding antagonists that display multiple binding modes. , 2002, Chemistry and Biology.

[9]  E. Toone,et al.  The cluster glycoside effect. , 2002, Chemical reviews.

[10]  Byron Goldstein,et al.  Analysis of cholera toxin-ganglioside interactions by flow cytometry. , 2002, Biochemistry.

[11]  P. Dub,et al.  A note on the problem of scattering from a single atomic plane and a stack of planes. Differences between the Ewald and other diffraction theories. , 2001 .

[12]  B. Sigurskjold,et al.  Exact analysis of competition ligand binding by displacement isothermal titration calorimetry. , 2000, Analytical biochemistry.

[13]  C. Verlinde,et al.  Using a Galactose Library for Exploration of a Novel Hydrophobic Pocket in the Receptor Binding Site of the Escherichia coliHeat-labile Enterotoxin* , 1999, The Journal of Biological Chemistry.

[14]  G. Whitesides,et al.  Nonstatistical binding of a protein to clustered carbohydrates. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[15]  J Angström,et al.  Binding of cholera toxin B-subunits to derivatives of the natural ganglioside receptor, GM1. , 1999, Journal of biochemistry.

[16]  Sandro Sonnino,et al.  Sugar Mimics: An Artificial Receptor for Cholera Toxin , 1999 .

[17]  George M Whitesides,et al.  Polyvalent Interactions in Biological Systems: Implications for Design and Use of Multivalent Ligands and Inhibitors. , 1998, Angewandte Chemie.

[18]  E A Merritt,et al.  The 1.25 A resolution refinement of the cholera toxin B-pentamer: evidence of peptide backbone strain at the receptor-binding site. , 1998, Journal of molecular biology.

[19]  C. Schengrund,et al.  Inhibition of the adherence of cholera toxin and the heat-labile enterotoxin of Escherichia coli to cell-surface GM1 by oligosaccharide-derivatized dendrimers. , 1998, Biochemical pharmacology.

[20]  K. Karlsson.  Meaning and therapeutic potential of microbial recognition of host glycoconjugates , 1998, Molecular microbiology.

[21]  S. Sonnino,et al.  Conformation of the oligosaccharide chain of G(M1) ganglioside in a carbohydrate-enriched surface. , 1998, Biophysical journal.

[22]  H. F. Fisher,et al.  Isoergonic cooperativity in glutamate dehydrogenase complexes: a new form of allostery. , 1997, Biochemistry.

[23]  P. Hajduk,et al.  Discovering High-Affinity Ligands for Proteins: SAR by NMR , 1996, Science.

[24]  J. McCann,et al.  Fluorescence analysis of galactose, lactose, and fucose interaction with the cholera toxin B subunit. , 1996, Biochemical and biophysical research communications.

[25]  R C Stevens,et al.  Cholera toxin binding affinity and specificity for gangliosides determined by surface plasmon resonance. , 1996, Biochemistry.

[26]  S. Homans,et al.  Solution dynamics of the oligosaccharide moiety of ganglioside GM1: Comparison of solution conformations with the bound state conformation in association with cholera toxin B‐pentamer , 1995, Journal of molecular recognition : JMR.

[27]  H. Wu,et al.  Fluorescence analysis of the interaction between ganglioside GM1-containing phospholipid vesicles and the B subunit of cholera toxin. , 1995, Biochimica et biophysica acta.

[28]  S. Teneberg,et al.  Delineation and comparison of ganglioside-binding epitopes for the toxins of Vibrio cholerae, Escherichia coli, and Clostridium tetani: evidence for overlapping epitopes. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[29]  K A Karlsson,et al.  On the role of the carboxyl group of sialic acid in binding of cholera toxin to the receptor glycosphingolipid, GM1. , 1994, Journal of biochemistry.

[30]  T. Sixma,et al.  Galactose‐binding site in Escherichia coli heat‐labile enterotoxin (LT) and cholera toxin (CT) , 1994, Molecular microbiology.

[31]  J. Martial,et al.  Crystal structure of cholera toxin B‐pentamer bound to receptor GM1 pentasaccharide , 1994, Protein science : a publication of the Protein Society.

[32]  M. Masserini,et al.  Fuc-GM1 ganglioside mimics the receptor function of GM1 for cholera toxin. , 1992, Biochemistry.

[33]  Domenico Acquotti,et al.  Three-dimensional structure of the oligosaccharide chain of GM1 ganglioside revealed by a distance-mapping procedure: a rotating and laboratory frame nuclear overhauser enhancement investigation of native glycolipid in dimethyl sulfoxide and in water-dodecylphosphocholine solutions , 1990 .

[34]  R. Bryant,et al.  Carbon-13 NMR of glycogen: hydration response studied by using solids methods. , 1989, Biochemistry.

[35]  J F Brandts,et al.  Rapid measurement of binding constants and heats of binding using a new titration calorimeter. , 1989, Analytical biochemistry.

[36]  K. Karlsson.  Animal glycosphingolipids as membrane attachment sites for bacteria. , 1989, Annual review of biochemistry.

[37]  R. Holmes,et al.  Comparison of the carbohydrate-binding specificities of cholera toxin and Escherichia coli heat-labile enterotoxins LTh-I, LT-IIa, and LT-IIb , 1988, Infection and immunity.

[38]  W. Jencks,et al.  On the attribution and additivity of binding energies. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[39]  G. Schwarzmann,et al.  Studies of the ligand binding to cholera toxin, II. The hydrophilic moiety of sialoglycolipids. , 1976, Hoppe-Seyler's Zeitschrift fur physiologische Chemie.

[40]  R. Brady,et al.  Preparation of radioactive Tay-Sachs ganglioside labeled in the sialic acid moiety. , 1970, Journal of lipid research.

[41]  R. Jeanloz,et al.  Acetohalogeno Derivatives and Glycosides of D-Galactosamine1 , 1958 .