The objective of this proposal is to elucidate the mechanical regulation of molecular interactions between selectins and glycoconjugate ligands, which mediate the first step of a multistep adhesion and signaling cascade for circulating leukocytes to attach to and migrate across vascular endothelium at sites of tissue injury or infection. These interactions are crucial, because their malfunction can result in a number of inflammatory and thrombotic disorders. Selectin-ligand interactions are regulated mechanically as they take place in the hydrodynamic environment of the circulation. Our hypothesis is that mechanical regulation of selectin-ligand binding kinetics results from specific atomic-level interactions that are dictated by the structures of these molecules. Force regulates bond dissociation by changing the energy landscape of these interactions and/or forming new interactions during force-induced fit and/or conformational changes, thereby eliciting slip and catch bonds. Transport regulates bond formation by influencing collision frequency and encounter time between interacting molecules, modulating the dependence of association kinetics on intrinsic docking. Since selectin-ligand binding kinetics determines cellular function under flow, including tethering, rolling, and aggregation, relatively minor structural differences that alter atomic-level interactions may have major consequences for physiology and pathology. This broad hypothesis will be tested in three integrated specific aims: 1) Define impact of structural variations in selectins and ligands on their interactions, 2) Define selectin-ligand interactions at the atomic level by molecular dynamics simulations, and 3) Define the consequences of ligand-specific alterations in L-selectin-dependent adhesion in vivo. The three specific aims combine experimental, theoretical and computational approaches, include in silico, in vitro, and in vivo studies, and span multiple scales from atomic-level mechanisms, single-cell and single-molecule kinetic/mechanics experiments, to whole animal physiology. This systematic study will clarify how the mechanical regulation of selectin-ligand binding kinetics enables leukocytes to adhere to blood vessel wall in the hydrodynamic environment of the circulation. Decoding how molecular structure determines this regulation will provide key insights into vascular physiology and pathology. As a result, the data may offer new therapeutic approaches to inhibiting pathological cell adhesion during inflammation and thrombosis.
We propose to elucidate the mechanical regulation of molecular interactions between selectins and glycoconjugate ligands, which mediate circulating white blood cells to adhere to vascular surface at sites of tissue injury or infection. This regulation is crucial because malfunction of selectin-ligand interactions can result in a number of inflammatory and thrombotic disorders. The data may offer new therapeutic approaches to inhibiting pathological cell adhesion during inflammation and thrombosis.
|Mehta-D'souza, Padmaja; Klopocki, Arkadiusz G; Oganesyan, Vaheh et al. (2017) Glycan Bound to the Selectin Low Affinity State Engages Glu-88 to Stabilize the High Affinity State under Force. J Biol Chem 292:2510-2518|
|Liu, Zhenghui; Yago, Tadayuki; Zhang, Nan et al. (2017) L-selectin mechanochemistry restricts neutrophil priming in vivo. Nat Commun 8:15196|
|Liu, Baoyu; Chen, Wei; Zhu, Cheng (2015) Molecular force spectroscopy on cells. Annu Rev Phys Chem 66:427-51|
|Chen, Yunfeng; Liu, Baoyu; Ju, Lining et al. (2015) Fluorescence Biomembrane Force Probe: Concurrent Quantitation of Receptor-ligand Kinetics and Binding-induced Intracellular Signaling on a Single Cell. J Vis Exp :e52975|
|Chingozha, Loice; Zhan, Mei; Zhu, Cheng et al. (2014) A generalizable, tunable microfluidic platform for delivering fast temporally varying chemical signals to probe single-cell response dynamics. Anal Chem 86:10138-47|
|Zhu, Cheng (2014) Mechanochemitry: a molecular biomechanics view of mechanosensing. Ann Biomed Eng 42:388-404|
|Lee, Cho-yin; Lou, Jizhong; Wen, Kuo-kuang et al. (2013) Actin depolymerization under force is governed by lysine 113:glutamic acid 195-mediated catch-slip bonds. Proc Natl Acad Sci U S A 110:5022-7|
|Zhang, Yan; Jiang, Ning; Zarnitsyna, Veronika I et al. (2013) P-selectin glycoprotein ligand-1 forms dimeric interactions with E-selectin but monomeric interactions with L-selectin on cell surfaces. PLoS One 8:e57202|
|Xiao, Botao; Tong, Chunfang; Jia, Xiaoling et al. (2012) Tyrosine replacement of PSGL-1 reduces association kinetics with P- and L-selectin on the cell membrane. Biophys J 103:777-85|
|Zarnitsyna, Veronika I; Zhu, Cheng (2011) Adhesion frequency assay for in situ kinetics analysis of cross-junctional molecular interactions at the cell-cell interface. J Vis Exp :e3519|
Showing the most recent 10 out of 14 publications