Intein-mediated ligation and cyclization of expressed proteins.
Protein splicing is a posttranslational processing event that releases an internal protein sequence from a protein precursor. During the splicing process the internal protein sequence, termed an intein, embedded in the protein precursor self-catalyzes its excision and the ligation of the flanking protein regions, termed exteins. The dissection of the splicing pathway, which involves the precise cleavage and formation of peptide bonds, and the identification of key catalytic residues at the splice junctions have led to the modulation of the protein splicing process as a protein engineering tool. Novel strategies have been developed to use intein-catalyzed reactions for the production and manipulation of proteins and peptides. These new approaches have broken down the size limitation barrier of chemical synthetic methods and are less technically demanding. The purpose of this article is to describe how to use self-splicing inteins in protein semisynthesis and backbone cyclization. The first two sections of the article provide a brief review of the distinct chemical steps that underlie protein splicing and intein enabled technology.
Xu
M Q
MQ
New England Biolabs, Inc., 32 Tozer Road, Beverly, Massachusetts 01915, USA.
Evans
T C
TC
D001426:Bacterial Proteins / Q000378:metabolism; D017931:DNA Primers / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005822:Genetic Vectors / Q000235:genetics; D066298:In Vitro Techniques; D015080:Mesna / Q000737:chemistry; D009687:Nuclear Proteins / Q000378:metabolism; D010456:Peptides, Cyclic / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D011506:Proteins / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D020040:Trans-Splicing
D001426:Bacterial Proteins; D017931:DNA Primers; D009687:Nuclear Proteins; D010456:Peptides, Cyclic; D011506:Proteins; D011993:Recombinant Fusion Proteins; C109277:TSSC3 protein; D015080:Mesna