scholarly journals Construction and Quantitation of a Selectable Protein Splicing Sensor Using Gibson Assembly and Spot Titers

2021 ◽  
Vol 1 (3) ◽  
Author(s):  
Daniel Woods ◽  
Danielle S. LeSassier ◽  
Ikechukwu Egbunam ◽  
Christopher W. Lennon
Keyword(s):  
Protein Splicing ◽  
Gibson Assembly

scholarly journals Structure of the branched intermediate in protein splicing

2014 ◽  
Vol 111 (23) ◽  
pp. 8422-8427 ◽  
Author(s):  
Z. Liu ◽  
S. Frutos ◽  
M. J. Bick ◽  
M. Vila-Perello ◽  
G. T. Debelouchina ◽  
...  
Keyword(s):  
Protein Splicing

scholarly journals Biological Applications of Protein Splicing

Cell ◽  
2010 ◽  
Vol 143 (2) ◽  
pp. 191-200 ◽  
Author(s):  
Miquel Vila-Perelló ◽  
Tom W. Muir
Keyword(s):  
Protein Splicing ◽  
Biological Applications

scholarly journals Ultrafast Protein Splicing is Common among Cyanobacterial Split Inteins: Implications for Protein Engineering

2012 ◽  
Vol 134 (28) ◽  
pp. 11338-11341 ◽  
Author(s):  
Neel H. Shah ◽  
Geoffrey P. Dann ◽  
Miquel Vila-Perelló ◽  
Zhihua Liu ◽  
Tom W. Muir
Keyword(s):  
Protein Engineering ◽  
Protein Splicing

scholarly journals Faster Protein Splicing with theNostoc punctiformeDnaE Intein Using Non-native Extein Residues

2013 ◽  
Vol 288 (9) ◽  
pp. 6202-6211 ◽  
Author(s):  
Manoj Cheriyan ◽  
Chandra Sekhar Pedamallu ◽  
Kazuo Tori ◽  
Francine Perler
Keyword(s):  
Protein Splicing

Protein splicing of a non‐canonical Clostridium thermocellum intein with N‐terminal Gln

2006 ◽  
Vol 20 (5) ◽  
Author(s):  
Taryn L Powers ◽  
Matthew J. Drago ◽  
Deirdre M. Dorval ◽  
Katherine R. Connor ◽  
Kenneth V. Mills
Keyword(s):  
Clostridium Thermocellum ◽  
Protein Splicing

scholarly journals Amino Acid Residues in Both the Protein Splicing and Endonuclease Domains of the PI-SceI Intein Mediate DNA Binding

1998 ◽  
Vol 273 (8) ◽  
pp. 4607-4615 ◽  
Author(s):  
Zening He ◽  
Michael Crist ◽  
Hsiao-ching Yen ◽  
Xiaoqun Duan ◽  
Florante A. Quiocho ◽  
...  
Keyword(s):  
Amino Acid ◽  
Dna Binding ◽  
Protein Splicing ◽  
Amino Acid Residues

scholarly journals Protein Splicing Mechanisms and Applications

IUBMB Life ◽  
2005 ◽  
Vol 57 (7) ◽  
pp. 469-476 ◽  
Author(s):  
Francine Perler
Keyword(s):  
Protein Splicing

Rational design of an improved photo-activatable intein for the production of head-to-tail cyclized peptides

2019 ◽  
Vol 400 (3) ◽  
pp. 417-427 ◽  
Author(s):  
Jana K. Böcker ◽  
Wolfgang Dörner ◽  
Henning D. Mootz
Keyword(s):  
Rational Design ◽  
Cyclic Peptide ◽  
Pathway Engineering ◽  
Protein Splicing ◽  
Peptide Formation ◽  
Split Intein ◽  
Individual Reaction ◽  
C And N ◽  
Important Position ◽  
Peptide Product

Abstract Head-to-tail cyclization of genetically encoded peptides and proteins can be achieved with the split intein circular ligation of peptides and proteins (SICLOPPS) method by inserting the desired polypeptide between the C- and N-terminal fragments of a split intein. To prevent the intramolecular protein splicing reaction from spontaneously occurring upon folding of the intein domain, we have previously rendered this process light-dependent in a photo-controllable variant of the M86 intein, using genetically encoded ortho-nitrobenzyltyrosine at a structurally important position. Here, we report improvements on this photo-intein with regard to expression yields and rate of cyclic peptide formation. The temporally defined photo-activation of the purified stable intein precursor enabled a kinetic analysis that identified the final resolution of the branched intermediate as the rate-determining individual reaction of the three steps catalyzed by the intein. With this knowledge, we prepared an R143H mutant with a block F histidine residue. This histidine is conserved in most inteins and helps catalyze the third step of succinimide formation. The engineered intein formed the cyclic peptide product up to 3-fold faster within the first 15 min after irradiation, underlining the potential of protein splicing pathway engineering. The broader utility of the intein was also shown by formation of the 14-mer sunflower trypsin inhibitor 1.

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