Premium
Determining the activation barrier and pH‐dependence of each step of protein splicing
Author(s) -
Donahue Colleen A.,
York Daniel J.,
Dorval Deirdre M.,
Reitter Julie N.,
Mills Kenneth V.
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.520.6
Subject(s) - intein , protein splicing , rna splicing , chemistry , peptide bond , cleavage (geology) , thioester , computational biology , peptide , biochemistry , combinatorial chemistry , microbiology and biotechnology , gene , biology , enzyme , paleontology , rna , fracture (geology)
Protein splicing is a post‐translational event by which an intervening polypeptide, the intein, facilitates its own excision from the flanking polypeptides, the exteins, and the ligation of the exteins. We are able to study the protein splicing mechanism of the Pyrococcus abyssi PolII intein in vitro , and can determine the rate of the overall splicing reaction as well as that of each step. We have estimated the activation barrier for the first step of the reaction, an amide to thioester rearrangement, as well as the third step of the reaction, cyclization of the side chain of Asn or Gln coupled to peptide bond cleavage. We will use these data to assess mechanistic models for the control of the order of the steps of splicing. This material is based upon work supported by the National Science Foundation under grant MCB‐0950245, the National Institutes of Health, and the Camille and Henry Dreyfus Foundation.