کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
10844056 1069300 2005 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
A fusion protein system for the recombinant production of short disulfide bond rich cystine knot peptides using barnase as a purification handle
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
A fusion protein system for the recombinant production of short disulfide bond rich cystine knot peptides using barnase as a purification handle
چکیده انگلیسی
The inhibitor cystine knot (ICK) structural motif has been found in several small proteins and peptides from plants, insects, marine molluscs, and also in human. It is defined by a triple β-sheet that is held together by three intramolecular disulfide bonds built by six conserved cysteine residues that generate a highly rigid and stable fold. We describe a procedure for the production of ICK peptides with correct disulfide bond connectivities via expression in Escherichia coli as fusion proteins with an enzymatically inactive variant of the Bacillus amyloliquefaciens RNAse barnase. Barnase directs the fused peptide to the culture medium and the fusion protein can be isolated by combined cation exchange/reverse-phase chromatography. The ICK peptides are released from the barnase expression and purification handle either by cyanogen bromide or by protease cleavage to give pure and correctly folded cystine knot peptides.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 39, Issue 1, January 2005, Pages 82-89
نویسندگان
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