From the Programa de Biologia Estrutural, Instituto de Bioquímica Médica, Instituto Nacional de Biologia Estrutural e Bioimagem, Centro Nacional de Ressonância Magnética Nuclear Jiri Jonas, Universidade Federal do Rio de Janeiro, 21941-902 Rio de Janeiro, RJ and.
J Biol Chem. 2013 Sep 27;288(39):28331-45. doi: 10.1074/jbc.M113.500926. Epub 2013 Aug 8.
c-Abl is a key regulator of cell signaling and is under strict control via intramolecular interactions. In this study, we address changes in the intramolecular dynamics coupling within the c-Abl regulatory unit by presenting its N-terminal segment (N-Cap) with an alternative function in the cell as c-Abl becomes activated. Using small angle x-ray scattering, nuclear magnetic resonance, and confocal microscopy, we demonstrate that the N-Cap and the Src homology (SH) 3 domain acquire μs-ms motions upon N-Cap association with the SH2-L domain, revealing a stabilizing synergy between these segments. The N-Cap-myristoyl tether likely triggers the protein to anchor to the membrane because of these flip-flop dynamics, which occur in the μs-ms time range. This segment not only presents the myristate during c-Abl inhibition but may also trigger protein localization inside the cell in a functional and stability-dependent mechanism that is lost in Bcr-Abl(+) cells, which underlie chronic myeloid leukemia. This loss of intramolecular dynamics and binding to the cellular membrane is a potential therapeutic target.
c-Abl 是细胞信号转导的关键调节因子,通过分子内相互作用受到严格控制。在这项研究中,我们通过呈现 c-Abl 激活时其 N 端片段(N-Cap)在细胞中具有替代功能,解决了 c-Abl 调节单元内分子内动力学偶联的变化。使用小角度 X 射线散射、核磁共振和共聚焦显微镜,我们证明 N-Cap 和Src 同源(SH)3 结构域在与 SH2-L 结构域结合时获得 μs-ms 运动,揭示了这些片段之间的稳定协同作用。N-Cap-豆蔻酰连接可能会触发蛋白质锚定在膜上,因为这些翻转动力学发生在 μs-ms 时间范围内。该片段不仅在 c-Abl 抑制期间呈现豆蔻酸,而且可能在功能性和稳定性依赖的机制中触发细胞内蛋白质定位,该机制在慢性髓细胞白血病的基础 Bcr-Abl(+) 细胞中丢失。这种分子内动力学的丧失和与细胞膜的结合是一个潜在的治疗靶点。