Institute of Molecular Genetics of the Czech Academy of Sciences
+ flagellum/cilium, + axoneme, + microtubule-based cytoskeleton, + kinetoplastid parasites, + mammalian cells, + advanced light and electron microscopy
We have developed a biochemical approach that facilitates the identification of protein constituents of structures at the tip of the trypanosome flagellum, such as the flagella connector and axonemal capping structure. Based on localization and functional studies, we obtained a detailed model of the structure of the flagella connector, an intriguing mobile membrane junction connecting two flagella of a single cell. This study makes the flagella connector the best understood flagellum tip structure. [pubmed] [doi]
We have imaged cells of Trypanosoma brucei and Leishmania major using expansion microscopy, which is based on the principle of physical expansion of samples. We optimized this super-resolution approach to study the cytoskeletal structures of these important parasites. We have shown that expansion microscopy is a very robust method that allows rapid localization of proteins in an entire cellular volume at high resolution. [pubmed] [doi]
We have developed an approach that allows tagging of proteins at their endogenous loci and their simultaneous inducible overexpression. This approach facilitates preparation of overexpression cell lines without the need for molecular cloning, which is difficult in the case of large or repetitive proteins. We validated the approach for a number of cytoskeletal proteins and demonstrated that an overexpression of proteins of up to 500 kDa in size can be achieved and the overexpression populations can serve as an excellent source for protein purification. [pubmed] [doi]
