LOCKING MECHANISM 3: HINGE CONFORMATION

OVERVIEW

This experiment aims to evaluate the ability of the locking mechanisms to lock hinges in open or closed conformations.

BACKGROUND INFORMATION

This experiment is a downstream experiment of single hinge assembly, purification, and locking mechanisms 1 and 2. Relevant background information for these steps can be found on their respective pages of the lab notebook.

After evaluating whether strands of the locking mechanism could recognize and displace their intended targets in isolation, we then assessed its ability to influence whether hinges adopted an open or closed conformation. We predicted that if the locking mechanism was able to influence which state hinges were in, open or closed, we would see a greater proportion of hinges in the close state when treated with the lock strand compared to no treatment and a lower proportion of closed hinges when treated with the key strand.

METHODS

H2 hinges were assembled and purified as described in Hinge Assembly and Hinge Extension. After hinges were purified they were then incubated with either lock at 30°C for 25 min. Hinges were also incubated with neither strand as a negative control.

RESULTS

A greater proportion of closed hinges were observed when the hinges were treated with lock compared to when treated with key or no treatment. When hinges were treated with key, this produced the lowest proportion of closed hinges.  

Details on these results will be published after the judging period in alignment with competition rules.

DISCUSSION

Discussion of results will be published after the judging period in alignment with competition rules.