Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Mini-intein Structures from Extremophiles Suggest a Strategy for Finding Novel Robust Inteins

Version 1 : Received: 7 May 2021 / Approved: 10 May 2021 / Online: 10 May 2021 (10:29:29 CEST)

A peer-reviewed article of this Preprint also exists.

Hiltunen, M.K.; Beyer, H.M.; Iwaï, H. Mini-Intein Structures from Extremophiles Suggest a Strategy for Finding Novel Robust Inteins. Microorganisms 2021, 9, 1226. Hiltunen, M.K.; Beyer, H.M.; Iwaï, H. Mini-Intein Structures from Extremophiles Suggest a Strategy for Finding Novel Robust Inteins. Microorganisms 2021, 9, 1226.

Abstract

Inteins are prevalent among extremophiles. Mini-inteins with robust splicing properties are of particular interest for biotechnological applications due to their small size. However, biochemical and structural characterization has still been limited to a small number of inteins, and only a few inteins serve as widely used tools in protein engineering approaches. We determined the crystal structure of a naturally-occurring Pol-II mini-intein from Pyrococcus horikoshii and compared it with two other natural mini-inteins from Pyrococcus horikoshii. Despite the similar sizes, the comparison revealed distinct differences in insertions and deletions, implying specific evolutionary pathways from distinct ancestral origins. Our studies suggest that sporadically distributed mini-inteins might be more promising for further protein engineering applications than the highly conserved mini-inteins. Structural investigations of more inteins could guide the shortest path to finding novel robust mini-inteins suitable for protein engineering purposes.

Keywords

protein splicing; intein; crystal structure; hyperthermophile; protein engineering

Subject

Biology and Life Sciences, Biochemistry and Molecular Biology

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