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Peptide bond cleavage adjacent to aspargine or glutamine
Author(s) -
Urbanski Laura M.,
Chin Stacy L.,
Reitter Julie N.,
Mills Kenneth V.
Publication year - 2013
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.27.1_supplement.789.6
Subject(s) - cleavage (geology) , chemistry , intein , fluorophore , peptide bond , peptide , stereochemistry , fluorescence , rna splicing , biochemistry , biology , gene , paleontology , rna , physics , quantum mechanics , fracture (geology)
Protein splicing is a post‐translational, self‐catalyzed process in which an intervening polypeptide, the intein, is self‐excised from the flanking polypeptides, or exteins. The third step of the splicing process is the cyclization of Asn or Gln coupled to peptide bond cleavage. We have studied this cyclization in model peptides with an N‐terminal Abz fluorophore and a C‐terminal Lys‐DNP quencher. Cyclization of Asn or Gln coupled to peptide bond cleavage results in fluorescence, which allows us to measure the rate of cyclization. The cyclization and cleavage can also be followed by HPLC and high resolution LC‐MS. Initial data suggest that the rate of Asn cyclization is significantly faster than Gln cyclization, that the rate of cyclization is increased by the influence of an upstream His, and that the cyclization is pH dependent. This material is based upon work supported by the National Science Foundation under grant MCB‐0950245 and by the Camille and Henry Dreyfus Foundation.