Mechanical properties of the P-selectin/PSGL-1 interaction

Marshall, Bryan T.; McEver, R.P.; Zhu, Cheng

doi:10.1109/iembs.2002.1134523

Cited by 3 publications

(6 citation statements)

References 2 publications

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“…Experimentally, various assays have been used to test the forced bond dissociation including atomic force microscopy or AFM [11][12][13][14][15] and optical trap or OT [16,17]. Using an AFM approach, for example, the existence of catch bond was first visualized from the forced bond lifetime [18], and the bond rupture force dependence on physical factors (approaching velocity and contact duration between the receptor and the ligand pair) [13] and on force history (reverse rate as a single value function of force) [15] were well determined for the forced dissociation of P-selectin-PSGL-1 bond.…”

Section: Introductionmentioning

confidence: 99%

Probe stiffness regulates receptor-ligand bond lifetime under force

Zhang

Lü

Long

2011

Sci. China Phys. Mech. Astron.

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Receptor-ligand bond dissociation under applied force is crucial to elucidate its biological functionality when the molecular bond is usually connected to a mechanical probe. While the impact of probe stiffness, k, on bond rupture force has recently attracted more and more attention, the mechanism of how it affects the bond lifetime, however, remains unclear. Here we quantified the dissociation lifetime of selectin-ligand bond using an optical trap assay with low stiffness ranging from 3.5×10 3 to 4.7×10 2 pN/nm. Our results indicated that bond lifetime yielded distinct distributions with different probe stiffness, implying the stochastic feature of bond dissociation. It was also found that the mean lifetime varied with probe stiffness and that the catch bond nature was visualized at k3.0×10 2 pN/nm. This work furthered the understanding of the forced dissociation of selectin-ligand bond at varied probe stiffness, which is physiologically relevant to the tethered rolling of leukocytes under blood flow.probe stiffness, bond lifetime, optical trap

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Section: Introductionmentioning

confidence: 99%

Probe stiffness regulates receptor-ligand bond lifetime under force

Zhang

Lü

Long

2011

Sci. China Phys. Mech. Astron.

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“…Both lipid (labeled by nitrobenzoxadiazole fluorescence) and P-selectin (labeled with G1) appeared uniformly distributed under confocal microscopy. However, lipid fluorescence, but not protein fluorescence, could quickly recover after photobleaching, indicating that the bilayer was continuous and that P-selectin was immobilized presumably by attaching its cytoplasmic tail to the PEI layer (26). This procedure ensures a uniform length of the extracellular domain outside the lipid bilayer for both P-and L-selectin.…”

Section: Irrelevance Of Polymer Elasticity Modelsmentioning

confidence: 99%

“…After 20 min incubation, the Petri dish was filled with 10 ml Hank's balanced salt solution with 1% Ig-free bovine serum albumin. The P-selectin and L-selectin bilayers so formed had molecular densities of a few hundred sites/mm 2 that resulted in infrequent binding (15-20%) to the (s)PSGL-1-, G1-, or DREG56-coated cantilever tips, as required for measuring singlebond interactions (26). The bilayers were immediately used in AFM experiments.…”

Section: Forming Selectin-reconstituted Bilayersmentioning

confidence: 99%

“…Our in-house-built AFM system and its functionalization with ligands and mAbs have been described previously (24)(25)(26). Briefly, a piezoelectric translator (PZT) (Poly Physik Instrument, Boston, MA) was used to actuate the cantilever (unsharpened gold-coated half-wafer cantilevers, Veeco Instruments, Woodbury, NY).…”

Section: Afm System and Cantilever Functionalizationmentioning

confidence: 99%

“…The AFM experiments were similar to those designed for measuring lifetimes of single molecular bonds, as previously described (24)(25)(26). Briefly, binding was enabled by actuating the ligand-or antibody-coated cantilever tip into contact with the selectin reconstituted bilayer.…”

Section: Determining Molecular Spring Constantmentioning

confidence: 99%

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Measuring Molecular Elasticity by Atomic Force Microscope Cantilever Fluctuations

Marshall

Sarangapani

et al. 2006

Biophysical Journal

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In single-molecule mechanics experiments the molecular elasticity is usually measured from the deformation in response to a controlled applied force, e.g., via an atomic force microscope cantilever. We have tested the validity of an alternative method based on a recently developed theory. The concept is to measure the change in thermal fluctuations of the cantilever tip with and without its coupling to a rigid surface via the molecule. The new method was demonstrated by its application to the elasticity measurements of L- and P-selectin complexed with P-selectin glycoprotein ligand-1 or their respective antibodies, which showed values comparable to those measured from the slope of the force-extension curve. L- and P-selectin were found to behave as nearly linear springs capable of sustaining large forces and strains without sudden unfolding. The measured spring constants of approximately 4 and approximately 1 pN/nm for L- and P-selectin, respectively, suggest that a physiological force of approximately 100 pN would result in an approximately 200% strain for the respective selectins.

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Shear‐enhanced binding of intestinal colonization factor antigen I of enterotoxigenic Escherichia coli

Tchesnokova¹,

McVeigh

Kidd

et al. 2010

Molecular Microbiology

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SummaryIn the intestine, enterotoxigenic Escherichia coli works against peristaltic forces, adhering to the epithelium via the colonization factor antigen I (CFA/I) fimbrial adhesin CfaE. The CfaE adhesin is similar in localization and tertiary (but not primary) structure to FimH, the type 1 fimbrial adhesin of uropathogenic E. coli, which shows shear-dependent binding to epithelial receptors by an allosteric catch-bond mechanism. Thus, we speculated that CfaE is also capable of shear-enhanced binding. Indeed, bovine erythrocytes coursing over immobilized CFA/I fimbriae in flow chambers exhibited low accumulation levels and fast rolling at low shear, but an 80-fold increase in accumulation and threefold decrease in rolling velocity at elevated shear. This effect was reversible and abolished by pre-incubation of fimbriae with anti-CfaE antibody. Erythrocytes bound to whole CfaE in the same shear-enhanced manner, but to CfaE adhesin domain in a shear-inhibitable fashion. Residue replacements designed to disrupt CfaE interdomain interaction decreased the shear dependency of adhesion and increased binding under static conditions to human intestinal epithelial cells. These findings indicate that close interaction between adhesive and anchoring pilin domains of CfaE keeps the former in a low-affinity state that toggles into a high-affinity state upon separation of two domains, all consistent with an allosteric catch-bond mechanism of CfaE binding.

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Mechanical properties of the P-selectin/PSGL-1 interaction

Cited by 3 publications

References 2 publications

Probe stiffness regulates receptor-ligand bond lifetime under force

Probe stiffness regulates receptor-ligand bond lifetime under force

Measuring Molecular Elasticity by Atomic Force Microscope Cantilever Fluctuations

Shear‐enhanced binding of intestinal colonization factor antigen I of enterotoxigenic Escherichia coli

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