Magic-Angle Spinning Frequencies beyond 300 kHz Are Necessary To Yield Maximum Sensitivity in Selectively Methyl Protonated Protein Samples in Solid-State NMR

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11310%2F18%3A10388243" target="_blank" >RIV/00216208:11310/18:10388243 - isvavai.cz</a>

Výsledek na webu

<a href="https://doi.org/10.1021/acs.jpcc.8b05600" target="_blank" >https://doi.org/10.1021/acs.jpcc.8b05600</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1021/acs.jpcc.8b05600" target="_blank" >10.1021/acs.jpcc.8b05600</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Magic-Angle Spinning Frequencies beyond 300 kHz Are Necessary To Yield Maximum Sensitivity in Selectively Methyl Protonated Protein Samples in Solid-State NMR

Popis výsledku v původním jazyce

In the last decade, proton detection in magic-angle spinning (MAS) solid-state NMR became a popular strategy for biomolecular structure determination. In particular, probe technology has experienced tremendous progress with smaller and smaller diameter rotors achieving ever higher MAS frequencies. MAS rotation frequencies beyond 100 kHz allow to observe and assign protons in fully protonated samples. In these experiments, resolution is however compromised as homogeneous proton-proton dipolar coupling interactions are not completely averaged out. Using a combination of experiments and simulations, we analyze the MAS frequency dependent intensities of the H-1,C-13 methyl correlation peaks of a selectively methyl protonated (CH3) microcrystalline sample of the chicken a-spectrin SH3 domain (alpha-SH3). Extensive simulations involving nine spins employing the program SIMPSON allow to predict the MAS frequency dependence of the proton intensities. The experimental results are used to validate the simulations. As quantitative measure, we determine the characteristic MAS frequency, which is necessary to obtain &gt;50% of the maximum achievable sensitivity. Our results show that this frequency is site-specific and strongly depends on the local methyl density. We find that the characteristic MAS frequency ranges from as low as 20 kHz up to 324 kHz with the average value of 135 +/- 88 kHz for this particular sample at a magnetic field strength of 11.7 T. Inclusion of side chain dynamics in the analysis reduces the average characteristic MAS frequency to 104 +/- 68 kHz within the range of 11-261 kHz. In case, &gt;80% of the maximum sensitivity shall be achieved, MAS rotation frequencies of 498 +/- 370 and 310 +/- 227 kHz are required with and without including side chains dynamics in the analysis, respectively.

Název v anglickém jazyce

Magic-Angle Spinning Frequencies beyond 300 kHz Are Necessary To Yield Maximum Sensitivity in Selectively Methyl Protonated Protein Samples in Solid-State NMR

Popis výsledku anglicky

In the last decade, proton detection in magic-angle spinning (MAS) solid-state NMR became a popular strategy for biomolecular structure determination. In particular, probe technology has experienced tremendous progress with smaller and smaller diameter rotors achieving ever higher MAS frequencies. MAS rotation frequencies beyond 100 kHz allow to observe and assign protons in fully protonated samples. In these experiments, resolution is however compromised as homogeneous proton-proton dipolar coupling interactions are not completely averaged out. Using a combination of experiments and simulations, we analyze the MAS frequency dependent intensities of the H-1,C-13 methyl correlation peaks of a selectively methyl protonated (CH3) microcrystalline sample of the chicken a-spectrin SH3 domain (alpha-SH3). Extensive simulations involving nine spins employing the program SIMPSON allow to predict the MAS frequency dependence of the proton intensities. The experimental results are used to validate the simulations. As quantitative measure, we determine the characteristic MAS frequency, which is necessary to obtain &gt;50% of the maximum achievable sensitivity. Our results show that this frequency is site-specific and strongly depends on the local methyl density. We find that the characteristic MAS frequency ranges from as low as 20 kHz up to 324 kHz with the average value of 135 +/- 88 kHz for this particular sample at a magnetic field strength of 11.7 T. Inclusion of side chain dynamics in the analysis reduces the average characteristic MAS frequency to 104 +/- 68 kHz within the range of 11-261 kHz. In case, &gt;80% of the maximum sensitivity shall be achieved, MAS rotation frequencies of 498 +/- 370 and 310 +/- 227 kHz are required with and without including side chains dynamics in the analysis, respectively.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10403 - Physical chemistry

Projekt

—

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2018

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Journal of Physical Chemistry C

ISSN

1932-7447

e-ISSN

—

Svazek periodika

122

Číslo periodika v rámci svazku

28

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

6

Strana od-do

16437-16442

Kód UT WoS článku

000439661000074

EID výsledku v databázi Scopus

2-s2.0-85048887010