Protein splicing: an analysis of the branched intermediate and its resolution by succinimide formation. |
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Authors: | M Q Xu D G Comb H Paulus C J Noren Y Shao and F B Perler |
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Institution: | New England Biolabs, Inc., Beverly, MA 01915. |
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Abstract: | Protein splicing involves the excision of an internal domain from a precursor protein and the ligation of the external domains so as to generate two new proteins. Study of this process has recently been facilitated by the isolation of a precursor and a branched intermediate from a thermophilic protein splicing element expressed in a foreign protein context. Two aspects of protein splicing are examined in this paper. We demonstrate a succinimide at the C-terminus of the spliced internal protein, implicating cyclization of asparagine in resolution of the branched intermediate, and we identify an alkali-labile bond in the branched intermediate. A revised protein splicing model based on these experimental results is presented. |
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