Solution structure of DnaE intein from Nostoc punctiforme: Structural basis for the design of a new split intein suitable for site-specific chemical modification

Oeemig, Jesper S.; Aranko, A. Sesilja; Djupsjöbacka, Janica; Heinämäki, Kimmo; Iwaï, Hideo

doi:10.1016/j.febslet.2009.03.058

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Volume 583, Issue 9 , Pages 1451-1456, 6 May 2009

Solution structure of DnaE intein from Nostoc punctiforme: Structural basis for the design of a new split intein suitable for site-specific chemical modification

Research Program in Structural Biology and Biophysics, Institute of Biotechnology, University of Helsinki, P.O. Box 65, Helsinki FIN-00014, Finland

Received 31 January 2009; received in revised form 13 March 2009; accepted 25 March 2009. published online 01 April 2009.

Edited by Hans Eklund

Abstract

Naturally split DnaE intein from Nostoc punctiforme (Npu) has robust protein trans-splicing activity and high tolerance of sequence variations at the splicing junctions. We determined the solution structure of a single chain variant of NpuDnaE intein by NMR spectroscopy. Based on the NMR structure and the backbone dynamics of the single chain NpuDnaE intein, we designed a functional split variant of the NpuDnaE intein having a short C-terminal half (C-intein) composed of six residues. In vivo and in vitro protein ligation of model proteins by the newly designed split intein were demonstrated.

Keywords: Intein, Protein splicing, NMR spectroscopy, Protein ligation, Nuclear spin relaxation, Chemical modification, Protein trans-splicing