What is it about?
These genes cloned by us encode two Rab7 GTPases in single-celled model eukaryote Paramecium octaurelia and display a very high homology to human counterparts. We used many methods such as: liquid chromatography-tandem mass spectrometry, quantitative Western blot, PCR,confocal/STED microscopy, in vivo electroporation, site-directed mutagenesis,in vitro glycosylation, quantitative assays with [P32] and [C14]UDPglucose,
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Why is it important?
Our data indicate that post-translational modifications (PTM) of Thr200 located in the hypervariable C-region of P.octaurelia Rab7b is crucial for the proper localization/function of this protein. Moreover, the two Rab7 paralogues differ also in another PTM: substantially more phosphorylated amino acid residues are in Rab7b than in Rab7a.
Perspectives
The two Paramecium octaurelia Rab7 paralogues differ from each other by five amino acids out of 206: four in the C-terminal hypervariable region. The fifth diverged amino acid residue in position 140 (Ser) in Rab7b and Ala in Rab7a is located in the region of a-helix. It was long-lasting but fascinating 'scientific trip' to uncover the secrets of nature.
Professor Elzbieta Wyroba
Nencki Institute of Experimental Biology
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This page is a summary of: Site‐directed mutagenesis, in vivo electroporation and mass spectrometry in search for determinants of the subcellular targeting of Rab7b paralogue in the model eukaryote, European Journal of Histochemistry, April 2016, PAGEPress Publications,
DOI: 10.4081/ejh.2016.2612.
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