Refereed journal article or data article (A1)

Novel alpha 2 beta 1 Integrin Inhibitors Reveal That Integrin Binding to Collagen under Shear Stress Conditions Does Not Require Receptor Preactivation

List of Authors: Nissinen L, Koivunen J, Käpylä J, Salmela M, Nieminen J, Jokinen J, Sipilä K, Pihlavisto M, Pentikäinen OT, Marjamäki A, Heino J


Publication year: 2012

Journal: Journal of Biological Chemistry


Journal acronym: J BIOL CHEM

Number in series: 53

Volume number: 287

Issue number: 53

Start page: 44694

End page: 44702

Number of pages: 9

ISSN: 0021-9258


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The interaction between alpha 2 beta 1 integrin (GPIa/IIa, VLA-2) and vascular collagen is one of the initiating events in thrombus formation. Here, we describe two structurally similar sulfonamide derivatives, BTT-3033 and BTT-3034, and show that, under static conditions, they have an almost identical effect on alpha 2-expressing CHO cell adhesion to collagen I, but only BTT-3033 blocks platelet attachment under flow (90 dynes/cm(2)). Differential scanning fluorimetry showed that both molecules bind to the alpha 2I domain of the recombinant alpha 2 subunit. To further study integrin binding mechanism(s) of the two sulfonamides, we created an alpha 2 Y285F mutant containing a substitution near the metal ion-dependent adhesion site motif in the alpha 2I domain. The action of BTT-3033, unlike that of BTT-3034, was dependent on Tyr-285. In static conditions BTT-3034, but not BTT3033, inhibited collagen binding by an alpha 2 variant carrying a conformationally activating E318W mutation. Conversely, in under flow conditions (90 dynes/cm(2)) BTT-3033, but not BTT-3034, inhibited collagen binding by an alpha 2 variant expressing E336A loss-of-function mutation. Thus, the binding sites for BTT-3033 and BTT-3034 are differentially available in distinct integrin conformations. Therefore, these sulfonamides can be used to study the biological role of different functional stages of alpha 2 beta 1. Furthermore, only the inhibitor that recognized the nonactivated conformation of alpha 2 beta 1 integrin under shear stress conditions effectively blocked platelet adhesion, suggesting that the initial interaction between integrin and collagen takes place prior to receptor activation.

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Last updated on 2022-07-04 at 16:33