Three-Dimensional Solid-State NMR Study of a Seven-Helical Integral Membrane Proton Pump—Structural Insights

Shi, Lichi; Ahmed, Mumdooh; Zhang, Wurong; Whited, Gregg; Brown, Leonid S.; Ladizhansky, Vladimir

doi:10.1016/j.jmb.2009.01.011

Cited by 145 publications

(181 citation statements)

References 71 publications

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“…The sequential resonance assignment is the basis of all further ssNMR spectroscopic studies (Pauli et al 2001;Schuetz et al 2010;Shi et al 2009;Sperling et al 2010). An important step was the development of efficient 15 N-13 C SPECIFIC-CP transfer which is widely used today (Baldus et al 1998).…”

Section: Introductionmentioning

confidence: 99%

BSH-CP based 3D solid-state NMR experiments for protein resonance assignment

et al. 2014

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We have recently presented band-selective homonuclear cross-polarization (BSH-CP) as an efficient method for CO-CA transfer in deuterated as well as protonated solid proteins. Here we show how the BSH-CP CO-CA transfer block can be incorporated in a set of threedimensional (3D) solid-state NMR (ssNMR) pulse schemes tailored for resonance assignment of proteins at high static magnetic fields and moderate magic-angle spinning rates. Due to the achieved excellent transfer efficiency of 33 % for BSH-CP, a complete set of 3D spectra needed for unambiguous resonance assignment could be rapidly recorded within 1 week for the model protein ubiquitin. Thus we expect that BSH-CP could replace the typically used CO-CA transfer schemes in well-established 3D ssNMR approaches for resonance assignment of solid biomolecules.

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Section: Introductionmentioning

confidence: 99%

BSH-CP based 3D solid-state NMR experiments for protein resonance assignment

et al. 2014

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“…Ideally, the location of the splicing site should have only minimal effects on protein structure or function. We therefore selected the BC loop because no distinct functional role was known so far, and only a small ␤ turn in contrast to the full ␤ sheet, as in bacteriorhodopsin, has been observed (18,19,30,31). Other regions, such as the EF loop, are known to affect color and protein stability (22,32), none of which has been reported for the BC loop.…”

Section: Resultsmentioning

confidence: 99%

“…Structural data of pR (18,19,31) indicate a ␤ turn motif in the BC loop. Although different studies suggest contradictory data for the exact position of this ␤ turn (Trp 83 -Gly 87 (19) and Gly 87 -Pro 90 (18)), the introduced mutations are in or next to the turn, respectively.…”

Section: Data In the Context Of Folding And Assembly Of ␣-Helicalmentioning

confidence: 99%

Assembling a Correctly Folded and Functional Heptahelical Membrane Protein by Protein Trans-splicing

Mehler¹,

Eckert

Busche³

et al. 2015

Journal of Biological Chemistry

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Background: Protein trans-splicing as a molecular design tool has been demonstrated for soluble but not yet for membrane proteins. Results: Two separate polypeptides have been spliced in vivo, yielding correctly folded and functional proteorhodopsin. Conclusion: Trans-splicing of ␣-helical membrane proteins under native conditions is possible. Significance: Our findings are important for the folding, assembly, and engineering of membrane proteins.

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“…[55,63] Außerdem konnten neue Proteinfamilien mittels FK-NMR-Spektroskopie untersucht werden. [118] Die Experimente zeigten, dass das Protein bereits im Grundzustand in unterschiedlichen Konformationen vorliegt.…”

Section: Membranproteineunclassified

Festkörper‐NMR‐Spektroskopie an komplexen Biomolekülen

2010

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Biomolekulare Anwendungen der NMR‐Spektroskopie werden oft mit löslichen Molekülen oder Magnetresonanztomographie in Verbindung gebracht. Seit Ende der 70er Jahre liefert darüber hinaus die Festkörper‐NMR‐Spektroskopie (FK‐NMR) auf atomarer Ebene Einblicke in komplexe biomolekulare Systeme von Lipid‐Doppelschichten bis hin zu Biomaterialien. Im letzten Jahrzehnt haben Fortschritte im Bereich der NMR‐Spektroskopie, der Biophysik und der Molekularbiologie das Repertoire der FK‐NMR‐Spektroskopie für biomolekulare Studien erheblich erweitert. Dieser Aufsatz behandelt neueste Ansätze und deren methodische Herausforderungen und diskutiert Fortschritte bei der Anwendung der FK‐NMR‐Spektroskopie im Grenzgebiet von Struktur‐ und Zellbiologie.

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Three-Dimensional Solid-State NMR Study of a Seven-Helical Integral Membrane Proton Pump—Structural Insights

Cited by 145 publications

References 71 publications

BSH-CP based 3D solid-state NMR experiments for protein resonance assignment

BSH-CP based 3D solid-state NMR experiments for protein resonance assignment

Assembling a Correctly Folded and Functional Heptahelical Membrane Protein by Protein Trans-splicing

Festkörper‐NMR‐Spektroskopie an komplexen Biomolekülen

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