{"publisher":"Springer Nature","date_published":"2012-08-23T00:00:00Z","type":"journal_article","article_type":"original","publication_identifier":{"issn":["0925-2738","1573-5001"]},"_id":"8463","page":"155-168","year":"2012","publication_status":"published","status":"public","author":[{"first_name":"Sam","last_name":"Asami","full_name":"Asami, Sam"},{"full_name":"Szekely, Kathrin","last_name":"Szekely","first_name":"Kathrin"},{"first_name":"Paul","full_name":"Schanda, Paul","orcid":"0000-0002-9350-7606","last_name":"Schanda","id":"7B541462-FAF6-11E9-A490-E8DFE5697425"},{"first_name":"Beat H.","last_name":"Meier","full_name":"Meier, Beat H."},{"last_name":"Reif","full_name":"Reif, Bernd","first_name":"Bernd"}],"intvolume":"        54","article_processing_charge":"No","abstract":[{"lang":"eng","text":"The 1H dipolar network, which is the major obstacle for applying proton detection in the solid-state, can be reduced by deuteration, employing the RAP (Reduced Adjoining Protonation) labeling scheme, which yields random protonation at non-exchangeable sites. We present here a systematic study on the optimal degree of random sidechain protonation in RAP samples as a function of the MAS (magic angle spinning) frequency. In particular, we compare 1H sensitivity and linewidth of a microcrystalline protein, the SH3 domain of chicken α-spectrin, for samples, prepared with 5–25 % H2O in the E. coli growth medium, in the MAS frequency range of 20–60 kHz. At an external field of 19.96 T (850 MHz), we find that using a proton concentration between 15 and 25 % in the M9 medium yields the best compromise in terms of sensitivity and resolution, with an achievable average 1H linewidth on the order of 40–50 Hz. Comparing sensitivities at a MAS frequency of 60 versus 20 kHz, a gain in sensitivity by a factor of 4–4.5 is observed in INEPT-based 1H detected 1D 1H,13C correlation experiments. In total, we find that spectra recorded with a 1.3 mm rotor at 60 kHz have almost the same sensitivity as spectra recorded with a fully packed 3.2 mm rotor at 20 kHz, even though ~20× less material is employed. The improved sensitivity is attributed to 1H line narrowing due to fast MAS and to the increased efficiency of the 1.3 mm coil."}],"extern":"1","citation":{"ista":"Asami S, Szekely K, Schanda P, Meier BH, Reif B. 2012. Optimal degree of protonation for 1H detection of aliphatic sites in randomly deuterated proteins as a function of the MAS frequency. Journal of Biomolecular NMR. 54(2), 155–168.","mla":"Asami, Sam, et al. “Optimal Degree of Protonation for 1H Detection of Aliphatic Sites in Randomly Deuterated Proteins as a Function of the MAS Frequency.” <i>Journal of Biomolecular NMR</i>, vol. 54, no. 2, Springer Nature, 2012, pp. 155–68, doi:<a href=\"https://doi.org/10.1007/s10858-012-9659-9\">10.1007/s10858-012-9659-9</a>.","chicago":"Asami, Sam, Kathrin Szekely, Paul Schanda, Beat H. Meier, and Bernd Reif. “Optimal Degree of Protonation for 1H Detection of Aliphatic Sites in Randomly Deuterated Proteins as a Function of the MAS Frequency.” <i>Journal of Biomolecular NMR</i>. Springer Nature, 2012. <a href=\"https://doi.org/10.1007/s10858-012-9659-9\">https://doi.org/10.1007/s10858-012-9659-9</a>.","apa":"Asami, S., Szekely, K., Schanda, P., Meier, B. H., &#38; Reif, B. (2012). Optimal degree of protonation for 1H detection of aliphatic sites in randomly deuterated proteins as a function of the MAS frequency. <i>Journal of Biomolecular NMR</i>. Springer Nature. <a href=\"https://doi.org/10.1007/s10858-012-9659-9\">https://doi.org/10.1007/s10858-012-9659-9</a>","ama":"Asami S, Szekely K, Schanda P, Meier BH, Reif B. Optimal degree of protonation for 1H detection of aliphatic sites in randomly deuterated proteins as a function of the MAS frequency. <i>Journal of Biomolecular NMR</i>. 2012;54(2):155-168. doi:<a href=\"https://doi.org/10.1007/s10858-012-9659-9\">10.1007/s10858-012-9659-9</a>","ieee":"S. Asami, K. Szekely, P. Schanda, B. H. Meier, and B. Reif, “Optimal degree of protonation for 1H detection of aliphatic sites in randomly deuterated proteins as a function of the MAS frequency,” <i>Journal of Biomolecular NMR</i>, vol. 54, no. 2. Springer Nature, pp. 155–168, 2012.","short":"S. Asami, K. Szekely, P. Schanda, B.H. Meier, B. Reif, Journal of Biomolecular NMR 54 (2012) 155–168."},"quality_controlled":"1","day":"23","publication":"Journal of Biomolecular NMR","issue":"2","title":"Optimal degree of protonation for 1H detection of aliphatic sites in randomly deuterated proteins as a function of the MAS frequency","month":"08","doi":"10.1007/s10858-012-9659-9","user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","date_updated":"2021-01-12T08:19:27Z","language":[{"iso":"eng"}],"volume":54,"date_created":"2020-09-18T10:09:18Z","oa_version":"None"}