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Old 12-29-2010, 04:04 PM
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Default Observing selected domains in multi-domain proteins via sortase-mediated ligation and NMR spectroscopy.

Observing selected domains in multi-domain proteins via sortase-mediated ligation and NMR spectroscopy.

Observing selected domains in multi-domain proteins via sortase-mediated ligation and NMR spectroscopy.

J Biomol NMR. 2010 Dec 29;

Authors: Refaei MA, Combs A, Kojetin DJ, Cavanagh J, Caperelli C, Rance M, Sapitro J, Tsang P

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 ~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.

PMID: 21188472 [PubMed - as supplied by publisher]



Source: PubMed
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