FHL3 (Four and a half LIM domain protein 3) is a protein consisting of four and a half LIM domains found mainly in skeletal muscle. LIM domains coordinate the zinc ion between four conserved amino acid residues and are crucial as interaction motifs between proteins. FHL3 influences the expression of certain genes, cell differentiation, and cytoskeleton formation and has an impact on cancer development through protein–protein interactions. It is one of the least characterized proteins of the FHL family. It interacts with transcription factors, cytoskeleton-related proteins, and various signaling molecules.
In the context of the thesis, we wanted to prepare a soluble form of FHL3 using the same procedure as was used to prepare a soluble form of FHL2. We inserted the transcript for our FHL3 protein into a linearised vector. To facilitate purification and isolation, we added to FHL3 fusion partners, hexahistidine tag (His$_6$) and sfGFP, which were cleaved during purification by the TEV protease. For expression, E. coli strain BL21[DE3] bacterial cells were used.
The protein was successfully expressed, purified, and isolated. The procedure used for the preparation of the soluble form of FHL2 proved to be effective for the preparation of the soluble form of FHL3. The prepared soluble form of FHL3 would be extremely useful for further research into its role in various cellular processes.
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