blog: membrane protein crystallography – is it still a fine art?

blog: membrane protein crystallography – is it still a fine art?

Membrane proteins serve a number of vital roles in the body, including oxygen production, transport of nutrients into cells, transport of water into and out of cells, removal of waste products, and allowing information to enter cells. Thirty percent of genes in the human genome encode for membrane proteins and 60% of drug targets are membrane proteins. Mutations or improper folding of membrane proteins are associated with many known diseases, such as heart disease, depression, obesity, and cancer. Dr. Isabel Moraes, Head of the Membrane Protein Laboratory (MPL) at Diamond Light Source (DLS), UK, has recently reviewed the methods used for the successful crystallisation of membrane proteins, summarised in our latest labCrystal journal.

Bottlenecks in membrane protein structure determination

In the last decade significant efforts have been made to improve the production and crystallisation of membrane proteins. Many of the crystallisation strategies have benefited from developments in automation and miniaturisation. For example, the use of liquid handling robots such as TTP Labtech’s mosquito® range has increased the number of potential crystallisation conditions that can be screened while at the same time reducing the amount of protein sample required.

In our new edition of labCrystal you can read about Dr. Moraes’ latest work on the crystallisation of anti-histamine drug targets. Dr. Moraes remarked, “In our lab, 99% of the solved membrane protein structures are from crystals set up with our TTP Labtech mosquito® LCP.”

Lipdic cubic phase technology miniaturised

In addition to the vapour diffusion method, membrane proteins are often crystallised in lipidic cubic phase (LCP). TTP Labtech’s mosquito LCP provides the combination of three important factors to Dr. Moraes’ team for successful crystallisation;

1) miniaturisation – volumes can be reduced to nanolitre ranges (as low as 25 nL)

2) speed – rapid dispensing of a 96-well LCP plate to about 4 mins

3) accuracy –  improved reproducibility with more accurate dispensing

Due to the fast set-up time, evaporation is reduced. However, TTP Labtech’s active humidity chamber is routinely used to avoid any possibility of drop evaporation.

Dr Moraes commented on the robustness of the mosquito LCP in a multi-user environment, “Because we operate as a user facility and have many different external scientists working in the lab, we really need a crystallisation machine that it is easy to work with and reliable….that’s our mosquito! I have been using several generations of the mosquito for more than 8 years and it has never let me down!”

If you would like to find out more about Dr. Moraes’ work or learn more about other successful crystallography projects that have benefitted from using TTP Labtech’s liquid handlers please download the latest labCrystal.

If you have any queries about TTP Labtech’s products please contact us at crystallography@ttplabtech.com.

Dr_Isabel_Moraes

Dr Isabel Moraes working with TTP Labtech’s mosquito LCP

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