title: Differences in zero-force and force-driven kinetics of ligand dissociation from beta-galactoside-specific proteins (plant and animal lectins, immunoglobulin G) monitored by plasmon resonance and dynamic single molecule force microscopy.
abstract: Protein-carbohydrate interactions are involved in diverse regulatory processes. To help understand the mechanics and kinetics of dissociation of receptor-ligand complexes, we have analyzed the separation of lactose and the N-glycan chains of asialofetuin (ASF) from three lectins and an immunoglobulin G fraction by surface plasmon resonance at zero force and by atomic force microscopy with variations of the external force. While the (AB)2 agglutinins from Ricinus communis (RCA) and Viscum album (VAA) show structural homology, the homodimeric galectin-1 from bovine heart (BHL) has no similarity to the two plant lectins except for sharing this monosaccharide specificity. The beta-galactoside-binding immunoglobulin G (IgG) fraction from human serum provides a further model system with distinct binding-site architecture. The k(off) constants for the two plant agglutinins were independent of the nature of the ligand at 1.1-1.3 x 10(-3) s(-1), whereas the geometry of ligand and binding site presentation affected this parameter for BHL (0.5 x 10(-3) s(-1) for lactose and 1 x 10(-3) s(-1) for ASF) and IgG (1.3 x 10(-3) s(-1) for lactose and 0.55 x 10(-3) s(-1) for ASF). When assessing comparatively the rupture forces at a loading rate of 3 nN/s with lactose as ligand, 34 +/- 6 pN (BHL), 36 +/- 4 pN (IgG), 47 +/- 7 pN (VAA), and 58 +/- 9 pN (RCA) were measured. For the same loading rate the rupture forces for the receptor-ASF interactions were found to be 37 +/- 3 pN (BHL), 43 +/- 5 pN (VAA), 45 +/- 6 pN (IgG), and 65 +/- 9 pN (RCA). The variation of the pulling velocity revealed in all cases a linear dependence between the rupture force and the natural logarithm of the loading rate. Performing probability density and Monte Carlo calculations, the potential barrier widths, which determine the inverse dynamic dependence with the rate of force elevation, increased from 4 A (RCA) and 7 A (VAA and IgG) to 10 A (BHL) for the receptor-lactose interactions. Presenting ASF as ligand potential widths of 4 A for RCA and IgG and 6 A for VAA and BHL were obtained. Since the dissociation kinetics at zero force apparently cannot predict the behavior in force-driven experiments, these results reveal new insights into biological functions. The dissociation kinetics under force helps to explain the difference in the toxic potency of VAA and RCA and points to a function of the galectin in cis-crosslinking and in transient trans-bridging.
authors.affiliation: Lehrstuhl für Angewandte Physik, Sektion Physik, Ludwig-Maximilians-Universität München, Germany.
authors.forename: W
authors.initials: W
authors.lastname: Dettmann
authors.forename: M
authors.initials: M
authors.lastname: Grandbois
authors.forename: S
authors.initials: S
authors.lastname: André
authors.forename: M
authors.initials: M
authors.lastname: Benoit
authors.forename: A K
authors.initials: AK
authors.lastname: Wehle
authors.forename: H
authors.initials: H
authors.lastname: Kaltner
authors.forename: H J
authors.initials: HJ
authors.lastname: Gabius
authors.forename: H E
authors.initials: HE
authors.lastname: Gaub
mesh_terms: D000818:Animals; D002417:Cattle; D005697:Galactosides / Q000737:chemistry / Q000378:metabolism*; D037483:Galectin 1; D006388:Hemagglutinins / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D007785:Lactose / Q000378:metabolism; D008024:Ligands; D018625:Microscopy, Atomic Force / Q000295:instrumentation / Q000379:methods*; D009010:Monte Carlo Method; D009206:Myocardium / Q000737:chemistry; D010940:Plant Proteins / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D020349:Surface Plasmon Resonance / Q000295:instrumentation / Q000379:methods*; D013816:Thermodynamics
chemical_list: D005697:Galactosides; D037483:Galectin 1; D006388:Hemagglutinins; D007074:Immunoglobulin G; D008024:Ligands; D010940:Plant Proteins; C007816:beta-galactoside; D007785:Lactose