Abstract
NMR spectroscopy has distinct advantages for providing insight into protein structures, but faces significant resolution challenges as protein size increases. To alleviate such resonance overlap issues, the ability to produce segmentally labeled proteins is beneficial. Here we show that the S. aureus transpeptidase sortase A can be used to catalyze the ligation of two separately expressed domains of the same protein, MecA (B. subtilis). The yield of purified, segmentally labeled MecA protein conjugate is approximately 40%. The resultant HSQC spectrum obtained from this domain-labeled conjugate demonstrates successful application of sortase A for segmental labeling of multi-domain proteins for solution NMR study.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Amino Acid Sequence
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Aminoacyltransferases / metabolism*
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Bacterial Proteins / chemistry
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Bacterial Proteins / metabolism*
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Cysteine Endopeptidases / metabolism*
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Magnetic Resonance Spectroscopy*
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Molecular Sequence Data
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Nitrogen Isotopes / chemistry
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Nuclear Magnetic Resonance, Biomolecular*
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Protein Structure, Tertiary
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Proteins / chemistry*
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Proteins / metabolism*
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / metabolism
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Staphylococcus aureus / enzymology
Substances
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Bacterial Proteins
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Nitrogen Isotopes
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Proteins
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Recombinant Fusion Proteins
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mecA protein, Bacillus subtilis
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Aminoacyltransferases
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sortase A
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Cysteine Endopeptidases