Journal
JOURNAL OF BIOMOLECULAR NMR
Volume 71, Issue 4, Pages 225-235Publisher
SPRINGER
DOI: 10.1007/s10858-018-0175-4
Keywords
Asparaginyl endopeptidase; Designed armadillo repeat protein; Segmental isotopic labeling; Protein trans-ligation; Protein NMR
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Funding
- Academy of Finland [137995, 277335]
- Biocenter Finland and Helsinki Institute of Life Science (HiLIFE)
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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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