+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

SAD phasing with in-house cu Ka radiation using barium as anomalous scatterer



SAD phasing with in-house cu Ka radiation using barium as anomalous scatterer



Indian Journal of Biochemistry and Biophysics 48(6): 399-405



Phasing of lysozyme crystals using co-crystallized barium ions was performed using single-wavelength anomalous diffraction (SAD) method using Cu Ka radiation with in-house source of data collection. As the ion binding sites vary with respect to the pH of the buffer during crystallization, the highly isomorphic forms of lysozyme crystals grown at acidic and alkaline pH were used for the study. Intrinsic sulphur anomalous signal was also utilized with anomalous signal from lower occupancy ions for phasing. The study showed that to solve the structure by SAD technique, 2.8-fold data redundancy was sufficient when barium was used as an anomalous marker in the in-house copper X-ray radiation source for data collection. Therefore, co-crystallization of proteins with barium containing salt can be a powerful tool for structure determination using lab source.

Please choose payment method:






(PDF emailed within 1 workday: $29.90)

Accession: 055629919

Download citation: RISBibTeXText

PMID: 22329242


Related references

Locating the anomalous scatterer substructures in halide and sulfur phasing. Acta Crystallographica. Section D, Biological Crystallography 59(Pt 1): 57-66, 2002

Locating the anomalous scatterer substructures in halide and sulfur phasing. Acta Crystallographica Section D Biological Crystallography 59(1): 57-66, January, 2003

Away from the edge: SAD phasing from the sulfur anomalous signal measured in-house with chromium radiation. Acta Crystallographica. Section D, Biological Crystallography 59(Pt 11): 1943-1957, 2003

Radiation-damage-induced phasing with anomalous scattering: substructure solution and phasing. Acta Crystallographica. Section D, Biological Crystallography 60(Pt 11): 1958-1963, 2004

In-house measurement of the sulfur anomalous signal and its use for phasing. Acta Crystallographica Section D Biological Crystallography 59(4): 688-696, April, 2003

Determining phases and anomalous-scattering models from the multiwavelength anomalous diffraction of native protein metal clusters. improved MAD phase error estimates and anomalous-scatterer positions. Acta Crystallographica. Section D, Biological Crystallography 53(Pt 1): 23-40, 1997

Away from the edge II: in-house Se-SAS phasing with chromium radiation. Acta Crystallographica. Section D, Biological Crystallography 61(Pt 7): 960-966, 2005

Can I solve my structure by SAD phasing? Anomalous signal in SAD phasing. Acta Crystallographica. Section D, Structural Biology 72(Pt 3): 346-358, 2016

In-house UV radiation-damage-induced phasing of selenomethionine-labeled protein structures. Journal of Structural Biology 181(2): 89-94, 2013

Comparison of phasing methods for sulfur-SAD using in-house chromium radiation: case studies for standard proteins and a 69 kDa protein. Acta Crystallographica. Section D, Biological Crystallography 61(Pt 11): 1533-1540, 2005

High-phasing-power lanthanide derivatives: taking advantage of ytterbium and lutetium for optimized anomalous diffraction experiments using synchrotron radiation. Acta Crystallographica. Section D, Biological Crystallography 59(Pt 10): 1877-1880, 2003

Structure of a fatty-acid-binding protein from Bacillus subtilis determined by sulfur-SAD phasing using in-house chromium radiation. Acta Crystallographica. Section D, Biological Crystallography 65(Pt 5): 440-448, 2009

Can I solve my structure by SAD phasing? Planning an experiment, scaling data and evaluating the useful anomalous correlation and anomalous signal. Acta Crystallographica. Section D, Structural Biology 72(Pt 3): 359-374, 2016

Determination of a novel structure by a combination of long-wavelength sulfur phasing and radiation-damage-induced phasing. Acta Crystallographica. Section D, Biological Crystallography 60(Pt 4): 686-695, 2004

30] Multiwavelength anomalous diffraction phasing of macromolecular structures: Analysis of MAD data as single isomorphous replacement with anomalous scattering data using the MADMRG program. Methods in Enzymology 276: 530-537, 1997