Lindhout Darrin A, Thiessen Angela, Schieve Dean, Sykes Brian D
Canadian Institutes of Health-Research Group in Protein Structure and Function, Department of Biochemistry, University of Alberta, Edmonton, Alberta, Canada T6G 2H7.
Protein Sci. 2003 Aug;12(8):1786-91. doi: 10.1110/ps.0376003.
Fusion protein constructs of the 56 amino acid globular protein GB-1 with various peptide sequences, coupled with the incorporation of a histidine tag for affinity purification, have generated high-yield fusion protein constructs. Methionine residues were inserted into the constructs to generate pure peptides following CNBr cleavage, yielding a system that is efficient and cost effective for isotopic labeling of peptides for NMR studies and other disciplines such as mass spectroscopy. Six peptides of varying sequences and hydrophobicities were expressed using this GB-1 fusion protein technique and produced soluble fusion protein constructs in all cases. The ability to easily express and purify recombinant peptides in high yields is applicable for biomedical research and has medicinal and pharmaceutical applications.
将56个氨基酸的球状蛋白GB-1与各种肽序列构建融合蛋白，并引入组氨酸标签用于亲和纯化，已产生了高产率的融合蛋白构建体。在构建体中插入甲硫氨酸残基，以便在溴化氰裂解后生成纯肽，从而得到一种高效且经济高效的系统，用于对肽进行同位素标记以供核磁共振研究及质谱等其他学科使用。利用这种GB-1融合蛋白技术表达了六种不同序列和疏水性的肽，并且在所有情况下均产生了可溶性融合蛋白构建体。能够轻松地高产表达和纯化重组肽，这适用于生物医学研究，并具有医学和制药应用。
