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A new on-axis multimode spectrometer for the macromolecular crystallography beamlines of the Swiss Light Source.

Owen RL, Pearson AR, Meents A, Boehler P, Thominet V, Schulze-Briese C - J Synchrotron Radiat (2009)

Bottom Line: In situ spectroscopic methods such as UV-Vis absorption and (resonance) Raman can provide this, and can also provide a means of detecting X-ray-induced changes.Here, preliminary results are introduced from an on-axis UV-Vis absorption and Raman multimode spectrometer currently being integrated into the beamline environment at X10SA of the Swiss Light Source.The continuing development of the spectrometer is also outlined.

View Article: PubMed Central - HTML - PubMed

Affiliation: Swiss Light Source, Paul Scherrer Institute, CH-5232 Villigen PSI, Switzerland.

ABSTRACT
X-ray crystallography at third-generation synchrotron sources permits tremendous insight into the three-dimensional structure of macromolecules. Additional information is, however, often required to aid the transition from structure to function. In situ spectroscopic methods such as UV-Vis absorption and (resonance) Raman can provide this, and can also provide a means of detecting X-ray-induced changes. Here, preliminary results are introduced from an on-axis UV-Vis absorption and Raman multimode spectrometer currently being integrated into the beamline environment at X10SA of the Swiss Light Source. The continuing development of the spectrometer is also outlined.

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Top: a charge transfer interaction between wild-type DsbB and ubiquinone induces a pink colouration in the quinone group. Inset bottom: in situ UV–Vis absorption spectroscopy of DsbAB crystals confirm the presence of the charge transfer interaction in the form of (bottom) a broad absorption peak centred at 510 nm.
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fig8: Top: a charge transfer interaction between wild-type DsbB and ubiquinone induces a pink colouration in the quinone group. Inset bottom: in situ UV–Vis absorption spectroscopy of DsbAB crystals confirm the presence of the charge transfer interaction in the form of (bottom) a broad absorption peak centred at 510 nm.

Mentions: Radiation effects were investigated in crystals of a second system, a ubiquinone binding E. coli membrane protein, DsbB, involved in disulfide bond formation (Malojcic et al., 2008 ▶). DsbB is known to induce a red shift in ubiquinone upon binding (Inaba et al., 2004 ▶) and a characteristic red-shifted visible peak appears upon ubiquinone binding within the DsbA–DsbB–ubiquinone (DsbAB-Q8) complex. Unlike heme-containing proteins in which the Soret band has a large molar extinction coefficient, this peak has a small extinction coefficient (∼4750 cm−1  M −1). However, despite the low intensity of this peak, it can be clearly observed in the SLS-MS with only 4 × 20 µs exposures (10 µm slits) (Fig. 8 ▶).


A new on-axis multimode spectrometer for the macromolecular crystallography beamlines of the Swiss Light Source.

Owen RL, Pearson AR, Meents A, Boehler P, Thominet V, Schulze-Briese C - J Synchrotron Radiat (2009)

Top: a charge transfer interaction between wild-type DsbB and ubiquinone induces a pink colouration in the quinone group. Inset bottom: in situ UV–Vis absorption spectroscopy of DsbAB crystals confirm the presence of the charge transfer interaction in the form of (bottom) a broad absorption peak centred at 510 nm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2651763&req=5

fig8: Top: a charge transfer interaction between wild-type DsbB and ubiquinone induces a pink colouration in the quinone group. Inset bottom: in situ UV–Vis absorption spectroscopy of DsbAB crystals confirm the presence of the charge transfer interaction in the form of (bottom) a broad absorption peak centred at 510 nm.
Mentions: Radiation effects were investigated in crystals of a second system, a ubiquinone binding E. coli membrane protein, DsbB, involved in disulfide bond formation (Malojcic et al., 2008 ▶). DsbB is known to induce a red shift in ubiquinone upon binding (Inaba et al., 2004 ▶) and a characteristic red-shifted visible peak appears upon ubiquinone binding within the DsbA–DsbB–ubiquinone (DsbAB-Q8) complex. Unlike heme-containing proteins in which the Soret band has a large molar extinction coefficient, this peak has a small extinction coefficient (∼4750 cm−1  M −1). However, despite the low intensity of this peak, it can be clearly observed in the SLS-MS with only 4 × 20 µs exposures (10 µm slits) (Fig. 8 ▶).

Bottom Line: In situ spectroscopic methods such as UV-Vis absorption and (resonance) Raman can provide this, and can also provide a means of detecting X-ray-induced changes.Here, preliminary results are introduced from an on-axis UV-Vis absorption and Raman multimode spectrometer currently being integrated into the beamline environment at X10SA of the Swiss Light Source.The continuing development of the spectrometer is also outlined.

View Article: PubMed Central - HTML - PubMed

Affiliation: Swiss Light Source, Paul Scherrer Institute, CH-5232 Villigen PSI, Switzerland.

ABSTRACT
X-ray crystallography at third-generation synchrotron sources permits tremendous insight into the three-dimensional structure of macromolecules. Additional information is, however, often required to aid the transition from structure to function. In situ spectroscopic methods such as UV-Vis absorption and (resonance) Raman can provide this, and can also provide a means of detecting X-ray-induced changes. Here, preliminary results are introduced from an on-axis UV-Vis absorption and Raman multimode spectrometer currently being integrated into the beamline environment at X10SA of the Swiss Light Source. The continuing development of the spectrometer is also outlined.

Show MeSH