Segmental isotopic labeling by asparaginyl endopeptidase-mediated protein ligation

被引：0
作者：
Kornelia M. Mikula
Luisa Krumwiede
Andreas Plückthun
Hideo Iwaï
机构：
[1] University of Helsinki,Research Program in Structural Biology and Biophysics, Institute of Biotechnology
[2] University of Zürich,Department of Biochemistry
来源：
Journal of Biomolecular NMR | 2018年 / 71卷
关键词：
Asparaginyl endopeptidase; Designed armadillo repeat protein; Segmental isotopic labeling; Protein ; -ligation; Protein NMR;
D O I：
暂无
中图分类号：
学科分类号：
摘要：
Segmental isotopic labeling can facilitate NMR studies of large proteins, multi-domain proteins, and proteins with repetitive sequences by alleviating NMR signal overlaps. Segmental isotopic labeling also allows us to investigate an individual domain in the context of a full-length protein by NMR. Several established methods are available for segmental isotopic labeling such as intein-mediated ligation, but each has specific requirements and limitations. Here, we report an enzymatic approach using bacterially produced asparagine endopeptidase from Oldenlandia affinis for segmental isotopic labeling of a protein with repetitive sequences, a designed armadillo repeat protein, by overcoming some of the shortcomings of enzymatic ligation for segmental isotopic labeling.
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页码：225 / 235
页数：10
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