Accurate Measurement of Alpha Proton Chemical Shifts of Excited Protein States by Relaxation Dispersion NMR Spectroscopy

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Lundström, Patrik,1971-University of Toronto, Departments of Biochemistry, Chemistry and Medical Genetics (författare)

Accurate Measurement of Alpha Proton Chemical Shifts of Excited Protein States by Relaxation Dispersion NMR Spectroscopy

Artikel/kapitelEngelska2009

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2009-01-16
American Chemical Society (ACS),2009
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Nummerbeteckningar

LIBRIS-ID:oai:DiVA.org:liu-51910
https://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-51910URI
https://doi.org/10.1021/ja807796aDOI

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Språk:engelska
Sammanfattning på:engelska

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Anmärkningar

Carr-Purcell-Meiboom-Gill relaxation dispersion NMR spectroscopy can provide detailed information about low populated, invisible states of protein molecules, including backbone chemical shifts of the invisible conformer and bond vector orientations that can be used as structural constraints. Notably, the measurement of H-1(alpha) chemical shifts in excited protein states has not been possible to date because, in the absence of suitable labeling, the homonuclear proton scalar coupling network in side chains of proteins leads to a significant degradation in the performance of proton-based relaxation dispersion experiments. Here we have overcome this problem through a labeling scheme in which proteins are prepared with U-H-2 glucose and 50% D2O/50% H2O that results in cleuteration levels of between 50-88% at the C-beta carbon. Effects from residual H-1(alpha)-H-1(beta) scalar couplings can be suppressed through a new NMR experiment that is presented here. The utility of the methodology is demonstrated on a ligand binding exchanging system and it is shown that H-1(alpha) chemical shifts extracted from dispersion profiles are, on average, accurate to 0.03 ppm, an order of magnitude better than they can be predicted from structure using a database approach. The ability to measure H-1(alpha) chemical shifts of invisible conformers is particularly important because such shifts are sensitive to both secondary and tertiary structure. Thus, the methodology presented is a valuable addition to a growing list of experiments for characterizing excited protein states that are difficult to study using the traditional techniques of structural biology.

Ämnesord och genrebeteckningar

NATURVETENSKAP Biologi Strukturbiologi hsv//swe
NATURAL SCIENCES Biological Sciences Structural Biology hsv//eng
NUCLEAR-MAGNETIC-RESONANCE; SH3 DOMAIN; SECONDARY STRUCTURE; SEQUENCE HOMOLOGY; DIPOLAR COUPLINGS; ADIABATIC PULSES; SPIN RELAXATION; C-ALPHA; DYNAMICS; C-13
Structural biology
Strukturbiologi

Biuppslag (personer, institutioner, konferenser, titlar ...)

Hansen, D. FlemmingUniversity of Toronto, Departments of Biochemistry, Chemistry and Medical Genetics (författare)
Vallurupalli, ParmodhUniversity of Toronto, Departments of Biochemistry, Chemistry and Medical Genetics (författare)
Kay, Lewis E.University of Toronto, Departments of Biochemistry, Chemistry and Medical Genetics (författare)
University of Toronto, Departments of Biochemistry, Chemistry and Medical Genetics (creator_code:org_t)

Sammanhörande titlar

Ingår i:Journal of the American Chemical Society: American Chemical Society (ACS)131:5, s. 1915-19260002-78631520-5126

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Av författaren/redakt...: Lundström, Patri ...; Hansen, D. Flemm ...; Vallurupalli, Pa ...; Kay, Lewis E.

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NATURVETENSKAP: NATURVETENSKAP; och Biologi; och Strukturbiologi

Artiklar i publikationen: Journal of the A ...

Av lärosätet: Linköpings universitet

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