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 Hint-domain [a]. The distance between the N- and C-terminal ends of the intein is comparable to 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 aim of our work was to create an intein-based chimeric protein for quick purification of the human growth hormone (HGH). We studied a number of chimeric proteins consisting of a short N-terminal peptide, the Mxe GyrA intein, and human growth hormone between them. Through the affinity purification mediated by specific tag on the intein, we created one-step purification protocol for Human Growth Hormone [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.
Publications:
Koonin EV, Starokadomskyy P. Are viruses alive? // Stud Hist Philos Biol Biomed Sci. 2016 Oct;59:125-134. PMID: 26965225
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
Starokadomskyy P.L., Okunev O.V., Dubey L.V. Utilizing of protein splicing phenomenon for optimization of obtaining and purification of the recombinant human growth hormone // Eur J Med Res., 2006, Oct. 8, vol.11 (17th European students Conference, 8-12 October 2006, Berlin) p. 128-129. (link)
Starokadomskyy P.L. Protein splicing // Molecular biology, 2007, vol. 41, N2, p. 278-293. PMID: 17514899
Starokadomskyy P., Dubey I.Ya., Okunev O.V., Irodov D.M. Construction of a Chimeric Intein-Containing Protein and the Search for Conditions for Its Cleavage // Cytology and Genetics, 2007, vol. 41, N2, p. 3-11. PMID: 17494337
