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|a Ravera, Enrico
|e author
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|a Massachusetts Institute of Technology. Department of Chemistry
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|a Francis Bitter Magnet Laboratory
|q (Massachusetts Institute of Technology)
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|a Corzilius, Bjorn
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|a Michaelis, Vladimir K.
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|a Griffin, Robert Guy
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|a Corzilius, Bjorn
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|a Michaelis, Vladimir K.
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|a Luchinat, Claudio
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|a Griffin, Robert Guy
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|a Bertini, Ivano
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|a DNP-Enhanced MAS NMR of Bovine Serum Albumin Sediments and Solutions
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|b American Chemical Society (ACS),
|c 2015-02-18T20:39:44Z.
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|z Get fulltext
|u http://hdl.handle.net/1721.1/94615
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|a Protein sedimentation sans cryoprotection is a new approach to magic angle spinning (MAS) and dynamic nuclear polarization (DNP) nuclear magnetic resonance (NMR) spectroscopy of proteins. It increases the sensitivity of the experiments by a factor of ∼4.5 in comparison to the conventional DNP sample preparation and circumvents intense background signals from the cryoprotectant. In this paper, we investigate sedimented samples and concentrated frozen solutions of natural abundance bovine serum albumin (BSA) in the absence of a glycerol-based cryoprotectant. We observe DNP signal enhancements of ε ∼ 66 at 140 GHz in a BSA pellet sedimented from an aqueous solution containing the biradical polarizing agent TOTAPOL and compare this with samples prepared using the conventional protocol (i.e., dissolution of BSA in a glycerol/water cryoprotecting mixture). The dependence of DNP parameters on the radical concentration points to the presence of an interaction between TOTAPOL and BSA, so much so that a frozen solution sans cryoprotectant still gives ε ∼ 50. We have studied the interaction of BSA with another biradical, SPIROPOL, that is more rigid than TOTAPOL and has been reported to give higher enhancements. SPIROPOL was also found to interact with BSA, and to give ε ∼ 26 close to its maximum achievable concentration. Under the same conditions, TOTAPOL gives ε ∼ 31, suggesting a lesser affinity of BSA for SPIROPOL with respect to TOTAPOL. Altogether, these results demonstrate that DNP is feasible in self-cryoprotecting samples.
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|a Deutsche Forschungsgemeinschaft (Research Fellowship CO 802/1-1)
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|a Natural Sciences and Engineering Research Council of Canada (NSERC Postdoctoral Fellowship)
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|a National Institutes of Health (U.S.) (grant EB-002804)
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|a National Institutes of Health (U.S.) (grant EB- 003151)
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|a National Institutes of Health (U.S.) (grant EB-002026)
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|a European Commission (contract East-NMR n. 228461)
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|a European Commission (contract Bio-NMR n. 261863)
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|a European Commission (COST action TD1103)
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|a Seventh Framework Programme (European Commission) (INSTRUCT (European FP7 e-Infrastructure grant, cont ract no. 211252))
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|a Italy. Ministero dell'istruzione, dell'università e della ricerca (MIUR PRIN (2009FAKHZT_001)
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|a Ente Cassa di risparmio di Firenze
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|a en_US
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|a Article
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|t Journal of Physical Chemistry B
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