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Post‐translational autoprocessing by an Oryza sativa hedgehog‐like domain
Author(s) -
Xega Kristina,
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.754.16
Subject(s) - hedgehog , gene , chemistry , biology , microbiology and biotechnology , genetics , stereochemistry
The Japanese wild rice Oryza sativa has a gene containing a hedgehog‐like autoprocessing domain. In order to determine if this domain can promote an amide to thioester rearrangement as with traditional hedgehog domains, we have amplified the gene by PCR and sub‐cloned it into an E. coli expression vector. We have over‐expressed the protein, purified it by affinity chromatography and analyzed the expressed protein by SDS‐PAGE, Western blot and MALDI‐TOF mass spectrometry. We are currently making a series of site‐directed mutations of the autoprocessing domain to explore the catalytic mechanism of the post‐translational cleavage. This material is based upon work supported by the National Science Foundation under grant MCB‐0950245 and by the Camille and Henry Dreyfus Foundation.