Revolution® System Pinpoints Enzyme Groups on Cell Membranes

An Andor Technology Revolution® laser spinning disk system, equipped with an ultrasensitive scientific digital camera, has helped German scientists understand how the same enzyme can regulate diverse cellular processes when grouped with other proteins. Their basic research could eventually benefit researchers developing anti-cancer treatments and drugs to prevent transplant rejection.

“Target of Rapamycin” (TOR) kinases, a type of enzyme, act on proteins that coordinate cell growth and division. To do this, they assemble together with other proteins into two groups or ‘complexes’ – TORC1 and TORC2. However, only TORC1 responds to rapamycin, an immune suppressant drug used to prevent organ transplant rejection. The role of TORC2 is less clear but it may help regulate the position of cell growth. Doris Berchtold and Tobias Walther from the Max Planck Institute of Biochemistry used an Andor Revolution® microscopy system with an Andor iXonEM+ 897 back-illuminated EMCCD to discover where within a cell TORC1 and TORC2 localise. This helps understand their different biological function and responsiveness to rapamycin.

They imaged fluorescent labels attached to components of the TOR complexes in yeast cells. Berchtold and Walther found TORC1 and TORC2 located in different, spatially-separated cellular compartments, with TORC2 acting in a previously unrecognised plasma membrane domain. There were few, very dynamic kinases in the cell.

“For ten years, scientists have tried to localize these kinases indirectly,” said Tobias Walther. “But, with this new generation of cameras, we can see single molecules in living cells – something which was impossible before.”

Walther gives credit to Andor’s Revolution® laser spinning disk system, which the company supplied to the lab’s specific requirements and includes photo-bleaching and activation capability.

“My lab does a lot of microscopy and for all of our applications, these new cameras have been a breakthrough. But it’s not just the cameras,” he says. “It’s also the coordination of all parts of the Revolution® system to achieve our goals that makes the difference - the little things that people don’t think much about until they use them.”