Determination of Kinetic Constants for the Interaction Between the Platelet Glycoprotein IIb-IIIa and Fibrinogen by Means of Surface Plasmon Resonance

Huber, Walter; Hurst, J.; Schlatter, Daniel; Barner, R.; Hübscher, Josef; Kouns, W C; Steiner, Beat

doi:10.1111/j.1432-1033.1995.tb20184.x

Cited by 57 publications

(33 citation statements)

References 42 publications

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“…These values are comparable with those determined by the same method for the interaction of integrins with their natural ligands such as between ␣ v ␤ 3 and osteopondin (K D ϭ 0.43 nM) (48), ␣ v ␤ 3 and the P1 fragment of laminin (K D ϭ 450 nM) (49), or ␣ IIb ␤ 3 and fibrinogen (K D ϭ 20 -70 nM) (50). For comparison, the affinity determined for the binding of the Fab-9 antibody to the ␣ IIb ␤ 3 integrin was also in the nanomolar range (51).…”

Section: Discussionsupporting

confidence: 81%

A Recombinant Chimeric Epidermal Growth Factor-like Module with High Binding Affinity for Integrins

Vella

Thielens

Bersch

et al. 2003

Journal of Biological Chemistry

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Integrins are cell surface receptors involved in numerous pathological processes such as metastasis invasion and abnormal angiogenesis. To target these receptors, the epidermal growth factor (EGF)-like domain of human complement protease C1r was used as a natural scaffold to design chimeric modules containing the RGD motif. Here we report a high yield bacterial expression system and its application to the production of two such modules, EGF-RGD and V2, the latter variant mimicking the RGD-containing domain of disintegrins. These modules were characterized chemically, and their biological activity was investigated by cellular assays using various Chinese hamster ovary cell lines expressing ␤ 1 and ␤ 3 integrins and by surface plasmon resonance spectroscopy. Remarkably, the modifications leading to the V2 variant had differential effects on the interaction with Targeting the appropriate cell type is a major challenge in cancer and cell therapy. Integrins, a family of cell receptors for plasma proteins, extracellular matrix proteins, and cell surface ligands (1-3), are potential targets for this purpose, because they are expressed by all cell types and are involved in many physiological functions including cell adhesion, proliferation, or differentiation, and pathological processes such as metastasis invasion, abnormal angiogenesis and vascularization, or thrombosis (4). In mammals, 18 ␣ and eight ␤ subunits assemble into 24 different non-covalent ␣␤ heterodimers that mediate bidirectional signals through the cell membrane. In response to cell activation, "inside-out" signals control the level of binding affinity for the ligand. In turn, ligand binding induces rearrangements in integrin structure that trigger "outside-in" signaling pathways (5). Extensive cross-talk also takes place between integrin and growth factor receptor signaling pathways (6, 7), influencing most integrin functions.The ␤ 1 integrin subset, comprising the major fibronectin receptor ␣ 5 ␤ 1 , is the most widely expressed. The ␤ 3 subset includes the fibrinogen receptor ␣ IIb ␤ 3 , present on platelets and megakaryocytes, and ␣ v ␤ 3 , originally described as the vitronectin receptor, which also binds a variety of matrix proteins including fibronectin and fibrinogen. Integrin ␣ v ␤ 3 is expressed on vascular cells during angiogenesis (8,9) and is also present in the adult on activated leukocytes, osteoclasts, and macrophages, where it participates in bone resorption and ingestion of apoptotic cells. The expression of various integrins has been shown to be deregulated in a number of cancer cells and invasive tumors and is thought to participate in the malignancy phenotypes (10 -14). Thus, a decreased expression of ␣ 5 ␤ 1 appears to increase the degree of tumorigenicity. In contrast, the level of ␣ v ␤ 3 is correlated with the survival and metastatic activity of tumor cells (10, 15) and has an important role in angiogenesis in tumors (11). Integrins, particularly ␣ v ␤ 3 , are therefore considered as appropriate targets for cancer therapy. Intrac...

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Section: Discussionsupporting

confidence: 81%

A Recombinant Chimeric Epidermal Growth Factor-like Module with High Binding Affinity for Integrins

Vella

Thielens

Bersch

et al. 2003

Journal of Biological Chemistry

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“…Our experiments suggest that the bond off‐rate may not be influenced by the Leu33Pro polymorphism, since both the Leu33 and Pro33 cells that adhered on fibrinogen at 25 s −1 could not be dislodged even after increasing the shear rate up to 1000 s −1 at the end of the perfusion (data not shown). Studies using surface plasmon resonance spectroscopy suggest that α IIb β 3 –fibrinogen binding consists of two consecutive processes: an initial fast reaction, with a reversible low affinity complex and a subsequent slower high affinity complex [28]. It is plausible that the combined presence of shear and the β 3 ‐helix disrupting residue proline could change the kinetic rates (and increase the on‐rate) of integrin α IIb β 3 to fibrinogen at one or both these steps.…”

Section: Discussionsupporting

confidence: 57%

Shear stress augments the enhanced adhesive phenotype of cells expressing the Pro33 isoform of integrin β₃

Vijayan¹,

Huang²,

Liu³

et al. 2003

FEBS Letters

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Adhesion of platelets to the exposed extracellular matrix proteins at sites of vascular injury is partly regulated by the local £uid shear stress. Because the Leu33Pro (Pl A ) polymorphism of integrin L L 3 confers only a modest increase in adhesion under static conditions, we used CHO and 293 cells expressing the Leu33 or Pro33 isoform of L L 3 in £ow chamber experiments to test whether shear forces would alter the Pl A adhesive phenotype. We found that shear force augmented the Pro33-mediated enhanced adhesion to ¢brinogen. This Pro33-dependent enhancement was aspirin-sensitive and was also observed on immobilized von Willebrand factor and cryoprecipitate, but not ¢bronectin. Thus, shear stress enhances the adhesive phenotype of the Pro33 cells to multiple physiologic substrates. ß

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“…This apparent discrepancy is clearly alleviated if the arrests we detected reflected a transient binding state. Further, other studies made on noncovalent ligand-receptor interactions revealed such intermediate states (31)(32)(33). Therefore, our three-parameter model may be considered as the simplest 1 We are grateful to Prof. Evan Evans for pointing out the importance of the difference of the forces experienced by the sphere and the bond.…”

Section: The Distribution Of Arrest Durations Is Quantitatively Consimentioning

confidence: 70%

Measuring the Lifetime of Bonds Made between Surface-linked Molecules

Pierrès

Benoliel

Bongrand

1995

Journal of Biological Chemistry

105

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It is not well known how the kinetic constants of association between soluble receptors and ligands may be used to predict the behavior of these molecules when they are bound to cell surfaces. Spherical beads were coated with varying densities of anti-rabbit immunoglobulin monoclonal antibodies and driven along glass surfaces derivatized with rabbit anti-dinitrophenol. Particle motion was analyzed. The velocity, attachment frequency, and duration of binding events were determined on individual particles. It was found that i) beads exhibited frequent arrests lasting between a few tenths of a second and more than one minute; ii) when antibodies were diluted, the median arrest duration remained fairly constant (Ϸ1 s) whereas binding frequency varied as the first power of the antibody concentration, suggesting that most particle arrests were due to the formation of a single bond; iii) when the shear rate was increased 7-fold, the duration of transient binding events remained constant. The disruptive force exerted on attachment points was estimated to range between about 6 and 37 piconewtons; and iv) the distribution of arrest durations suggested that binding was not a monophasic reaction but involved at least one intermediate step. Therefore, transient binding events reflected the formation of unstable associations that are not detected with standard techniques.An obvious requirement for a molecular understanding of cell adhesion would be to obtain a precise knowledge of the rates of bond formation and dissociation between membraneassociated receptors and ligands. Indeed, it was recently emphasized that the outcome of an intercellular contact might be more dependent on the kinetics than the affinity of interaction between ligands and receptors (1-3). Thus, the capacity of adhesion molecules such as selectins to allow the rolling of leukocytes along endothelial cells in flowing blood was suggested to rely on a particularly high value of kinetic constants (3). Also, when a first bond occurred between a cell and another cell or surface, a critical parameter of adhesion is the ratio between the rates of dissociation of the first bond and formation of additional interactions (4).However, to our knowledge, no previously reported methodology allowed a direct measurement of the lifetime of interactions between particle-bound molecules (5). Tha et al. (6) used a travelling microtube to study the time and force dependence of rupture of antibody-mediated erythrocyte doublets. However, they did not study very transient attachments. Wattenbarger et al. (7) studied the adhesion of glycophorin-containing liposomes to a lectin-coated surface in shear flow. Although they studied the motion of individual particles, they did not present quantitative data on short-term arrests. Other experiments done with the parallel plate flow chamber yielded direct information on binding efficiency and binding strength rather than binding kinetics (8, 9). Also, Evans et al. (10) performed micromanipulation to determine the mechanical resistance of ...

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Determination of Kinetic Constants for the Interaction Between the Platelet Glycoprotein IIb-IIIa and Fibrinogen by Means of Surface Plasmon Resonance

Cited by 57 publications

References 42 publications

A Recombinant Chimeric Epidermal Growth Factor-like Module with High Binding Affinity for Integrins

A Recombinant Chimeric Epidermal Growth Factor-like Module with High Binding Affinity for Integrins

Shear stress augments the enhanced adhesive phenotype of cells expressing the Pro33 isoform of integrin β₃

Measuring the Lifetime of Bonds Made between Surface-linked Molecules

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Determination of Kinetic Constants for the Interaction Between the Platelet Glycoprotein IIb-IIIa and Fibrinogen by Means of Surface Plasmon Resonance

Cited by 57 publications

References 42 publications

A Recombinant Chimeric Epidermal Growth Factor-like Module with High Binding Affinity for Integrins

A Recombinant Chimeric Epidermal Growth Factor-like Module with High Binding Affinity for Integrins

Shear stress augments the enhanced adhesive phenotype of cells expressing the Pro33 isoform of integrin β3

Measuring the Lifetime of Bonds Made between Surface-linked Molecules

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Shear stress augments the enhanced adhesive phenotype of cells expressing the Pro33 isoform of integrin β₃