Abstract
HER2-directed chimeric antigen receptor (CAR) T cells have demonstrated robust in vivo cytotoxic activity against HER2-positive tumors. However, the structural basis of CAR activation and signal transduction across the membrane remains poorly defined due to the lack of full-length, high-resolution CAR structures. Here, we used homology modeling and all-atom molecular dynamics simulations totaling 37.7 µs to probe the structural dynamics of a full-length anti-HER2 CAR embedded in an explicit POPC membrane, sampling both apo-form and antigen-bound (holo-form) states across three independent trajectories. The simulations reveal coordinated, antigen-dependent changes in the dynamics of the extracellular antigen-binding domain (AB) and the intracellular signaling domain (SI), coupled through the intrinsic disordered regions including hinge, transmembrane, and costimulatory domains (HI-TM-CS). In the apo state, AB exhibits broad extracellular conformational sampling consistent with antigen search, whereas intracellular SI sampling remains comparatively restricted. Upon antigen engagement, this dynamic coupling is altered in a manner reminiscent of an oscillating coupled pendulum, with reduced AB spatial sampling and increased intracellular SI mobility, consistent with a redistribution of domain dynamics that may facilitate accessibility of signaling motifs and downstream kinase engagement, like LCK phosphorylation. We refer to this antigen-dependent reversal in coupled domain dynamics as a Binding-Induced Domain Dynamics Switch (BIDDS). BIDDS differs from static safety on/off models of T cell receptor activation and is presented as a mechanistic hypothesis to guide future computational and experimental studies of CAR activation and related multi-domain receptor signaling, with potential relevance to other receptor tyrosine kinases including VEGFR, EGFR, and FGFR.