Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.
J Biol Chem. 2010 Nov 12;285(46):35967-78. doi: 10.1074/jbc.M110.155770. Epub 2010 Sep 6.
Binding of lymphocyte function-associated antigen-1 (LFA-1) to intercellular adhesion molecule-1 (ICAM-1) mediates leukocyte adhesion under force. Using a biomembrane force probe capable of measuring single bond interactions, we showed ICAM-1 binding to LFA-1 at different conformations, including the bent conformation with the lowest affinity. We quantify how force and conformations of LFA-1 regulate its kinetics with ICAM-1. At zero-force, on-rates were substantially changed by conditions that differentially favor a bent or extended LFA-1 with a closed or open headpiece; but off-rates were identical. With increasing force, LFA-1/ICAM-1 bond lifetimes (reciprocal off-rates) first increased (catch bonds) and then decreased (slip bonds). Three states with distinct off-rates were identified from lifetime distributions. Force shifted the associated fractions from the short- to intermediate- and long-lived states, producing catch bonds at low forces, but increased their off-rates exponentially, converting catch to slip bonds at high forces. An internal ligand antagonist that blocks pulling of the α(7)-helix suppressed the intermediate-/long-lived states and eliminated catch bonds, revealing an internal catch bond between the αA and βA domains. These results elucidate an allosteric mechanism for the mechanochemistry of LFA-1/ICAM-1 binding.
淋巴细胞功能相关抗原-1(LFA-1)与细胞间黏附分子-1(ICAM-1)的结合介导了白细胞在力的作用下的黏附。我们使用一种能够测量单键相互作用的生物膜力探针,展示了不同构象下 ICAM-1 与 LFA-1 的结合,包括亲和力最低的弯曲构象。我们量化了力和 LFA-1 的构象如何调节其与 ICAM-1 的动力学。在零力条件下,通过有利于 LFA-1 呈现弯曲或伸展、头基闭合或打开的条件,显著改变了进入速率;而离开速率则相同。随着力的增加,LFA-1/ICAM-1 键的寿命(倒数离开速率)首先增加(捕获键),然后减少(滑动键)。从寿命分布中确定了具有不同离开速率的三种状态。力将关联分数从短寿命状态转移到中寿命和长寿命状态,在低力下产生捕获键,但随着力的增加,其离开速率呈指数增加,将捕获键转换为滑动键。一种阻断α(7)-螺旋拉动的内部配体拮抗剂抑制了中/长寿命状态,并消除了捕获键,揭示了αA 和βA 结构域之间的内部捕获键。这些结果阐明了 LFA-1/ICAM-1 结合的机械化学的变构机制。