Trans-splicing inteins

Martin DD, Xu MQ, Evans TC Jr. Characterization of a naturally occurring trans-splicing intein from Synechocystis sp. PCC6803. Biochemistry 2001 40(5) 1393-1402. 

Fig. 1.5 A Natural protein splicing, B Trans splicing with a split intein. The two fragments can be prepared separately and reassembled in vitro to form an active intein domain for protein ligation. C Central segment labeling using two different split inteins.

Fig. 9.10. Ligation of two protein fragments by trans-splicing interns a) on the DNA level, the two fragments can be encoded in separate ORFs and are flanked by either a 3 -extension encoding the N-intein (N-fragment) or 5 -extension encoding the C-intein (C-fragment). b) Following transcription and translation, the intein domains dimerize and c) excise themselves, fusing the N- and C-fragment by a peptide backbone bond.

Evans, T. C. Jr., Martin, D., Kolly, R., et al. (2000) Protein trans-splicing and cyclization by a naturally split intein from the dnaE gene of Synechocystis species PCC6803. /. Biol. Chem. 275, 9091-9094. 

Shingledecker K, Jiang SQ, Paulus H. Molecular dissection of the Mycobacterium tuberculosis RecA intein design of a minimal intein and of a trans-splicing system involving two intein fragments. Gene 1998 207(2) 187-195. 

Wu H, Hu ZM, Liu XQ. Protein trans-splicing by a split intein encoded in a spht dnae gene of synechocystis sp. Pcc6803. Proc. Natl. Acad. Sci. U.S.A. 1998 95(16) 9226-9231. 

Stm W, Yang J, Liu XQ. Synthetic two-piece and three-piece spht inteins for protein trans-splicing. J. Biol. Chem. 2004 279(34) 35281-35286. 

Shi J, Muir TW. Development of a tandem protein trans-splicing system based on native and engineered split inteins. J. Am. Chem. Soc. 2005 127(17) 6198-6206. 

Kwon Y, Coleman MA, Camarero JA. Selective immobilization of proteins onto solid supports through spUt-intein-mediated protein trans-splicing. Angew. Chem/ Int. Ed. Engl. 2006 45(11) 1726-1729. 

Dhar T, Mootz HD (2011) Modification of transmembrane and GPI-anchored proteins on living cells by efficient protein trans-splicing using the Npu DnaE intein. Chem Commun 47 3063-3065... 

Figure 4 Mechanism of trans-protein splicing, (a) Initial association of the intein halves to form a functional intein. (b) Activation of the N-terminal splice-junction via an N-S acyl shift, (c) Formation of a branched intermediate upon transthioesterification. (d) Branch resolution and intein release by succinimide formation. Spontaneous S-N acyl rearrangement yields the processed product with a native peptide backbone.

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