کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1178336 962684 2007 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Preceding hydrophobic and β-branched amino acids attenuate splicing by the CnePRP8 intein
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Preceding hydrophobic and β-branched amino acids attenuate splicing by the CnePRP8 intein
چکیده انگلیسی

As the Cne PRP8 intein is active and exists in an essential gene of an important fungal pathogen, inhibitors of splicing and assays for intein activity are of interest. The self-splicing activity of Cne PRP8, the intein from the Prp8 gene of Cryptococcus neoformans, was assessed in different heterologous fusion proteins expressed in Escherichia coli. Placement of a putatively inactive variant of the intein adjacent to the α-complementation peptide abolished the peptide's ability to restore β-galactosidase activity, while an active variant allowed complementation. This α-complementation peptide therefore provides a facile assay of splicing which can be used to test potential inhibitors. When placed between two heterologous protein domains, splicing was impaired by a β-branched amino acid immediately preceding the intein, while splicing occurred only with a hydroxyl or thiol immediately following the intein. Both these assays sensitively report impairment of splicing and provide information on how context constrains the splicing ability of Cne PRP8.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics - Volume 1774, Issue 8, August 2007, Pages 995–1001
نویسندگان
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