In kinesin's mechanochemical cycle, ATP's binding to the nucleotide-free leading head is exquisitely gated so that futile hydrolysis is effectively avoided, Experiments show that, when both kinesin heads bind to a m...In kinesin's mechanochemical cycle, ATP's binding to the nucleotide-free leading head is exquisitely gated so that futile hydrolysis is effectively avoided, Experiments show that, when both kinesin heads bind to a microtubule~ ATP cannot bind to kinesin's leading head when the neck linker (NL) of this head has a backward orientation. How NL's backward orientation is maintained needs understanding on a structural basis. By using steered molecular dynamics and rrmtation simulations, we investigate the backward-pointing conformation of the leading head's NL under different inter-head tensions. We find that the NL cannot keep in a strict backward orientation solely by the inter-head tension. LYS325 (amino acid sequence in 2KIN) has an assistant locking function which locks the NL and β0 to the β-domain. This locking function has an enhanced positive cooperation with the inter-head tension. When the inter-head tension is weakened, this locking function can be broken, resulting in a loose backward orientation of the NL. The difference between the strict and loose backward orientation of the NL might be a crucial factor in the gating mechanism. These results are consistent with relevant experiments and proposals.展开更多
基金Supported by the National Natural Science Foundation of China under Grant Nos 90403007, 10975044 and 10975019, the Foundation of the Ministry of Personnel of China for Returned Scholars under Grant No MOP2006138, the Eundamental Research Funds for the Central University and the Key Subject Construction Project of Hebei Provincial Universities.
文摘In kinesin's mechanochemical cycle, ATP's binding to the nucleotide-free leading head is exquisitely gated so that futile hydrolysis is effectively avoided, Experiments show that, when both kinesin heads bind to a microtubule~ ATP cannot bind to kinesin's leading head when the neck linker (NL) of this head has a backward orientation. How NL's backward orientation is maintained needs understanding on a structural basis. By using steered molecular dynamics and rrmtation simulations, we investigate the backward-pointing conformation of the leading head's NL under different inter-head tensions. We find that the NL cannot keep in a strict backward orientation solely by the inter-head tension. LYS325 (amino acid sequence in 2KIN) has an assistant locking function which locks the NL and β0 to the β-domain. This locking function has an enhanced positive cooperation with the inter-head tension. When the inter-head tension is weakened, this locking function can be broken, resulting in a loose backward orientation of the NL. The difference between the strict and loose backward orientation of the NL might be a crucial factor in the gating mechanism. These results are consistent with relevant experiments and proposals.