summary:Protein splicing is a post-translational process that results in excision of an internal protein region (intein) and ligation of its flanking sequences (exteins). Protein splicing does not require cofactors of host's enzymes: it is catalyzed by itself through the internal domain (Hint-domain) [a]. It can be explained by tertiary structure of intein. The distance between the N and C ends of the intein is comparable with the length of a covalent bond (about 1.4 Å). Hint-domain plays an important role in autocatalytic cleavage of the intein and subsequent ligation of the exteins. Interestingly, that the Hint-domain catalytic center structurally is similar to the active center of serine protease.
The main aims of our work was to create intein-based chimeric protein for quick purification of the human growth hormone (HGH). I made a chimeric protein consisting of a short N-terminal peptide, the Mxe GyrA intein, and human growth hormone between them. I did quick affinity purification by the intein, and then induce self-cleavage of the intein, that releases human growth hormone in a pure buffer [b, c].
LOCATION: Institute of molecular biology and genetics NAS of Ukraine, Kiev
Obtained degree and awards:
Second place for the presentation "Optimization of recombinant protein obtaining and purification using protein splicing" on Conference of Young Scientists "Actual problems of biochemistry and biotechnology - 2006", Kiev, Ukraine;
First place award in the category of "Best cycle of scientific publication in field of biotechnology" given by Institute of Molecular biology and Genetics of NAS of Ukraine
Starokadomskyy P.L., Okunev O.V. , Irodov D.M., Kordium V.A.Utilization of Protein Splicing for Purification of the Human Growth Hormone // Molecular biology (Moscow), 2008, Vol. 42, No. 6, pp. 966-972. PMID: 19140330