PIN5 and PIN8 the Yin and Yang of membrane topology

“Mapping the membrane orientation of auxin homeostasis regulators PIN5 and PIN8 in Arabidopsis thaliana root cells reveals their divergent topology” is on-line (and in pdf).

Big congratulations to Yewubnesh and the team Vendula, Nikola, Veronika, Marta and Marek. We are very happy to contribute to the very active research field of Endoplasmic Reticulum PIN auxin carriers. Yewubnesh has performed formidable cell-biology work and, in the process, provided a tool-kit to study the membrane topology of ER proteins using live and immuno-staining techniques. The microscopy-based part of the research would not be possible without CEITEC CELLIM.

I do love the cell biology approaches also those utilising digitonin but I am happy that we were able to benefit from the biochemistry expertise of Veronika Bilanovicova and supplement (Fig. S4) the microscopy-based techniques by a western-visualised trypsin digestion of apoplast-facing protein parts, including the hydrophilic loop of the ectopically PM localised PIN8.

My thanks go also Vendula, Nikola, Marta and Marek who helped with the digitonin protocols optimisation and the last but critical steps of PIN8 and PIN5 transgenic lines phenotyping. Here especially Vendula help was very timely.