Structure determination by multiple-wavelength anomalous dispersion (MAD) at the Pr L_IIIedge

Abstract

The use of longer X-ray wavelengths in macromolecular crystallography has grown significantly over the past few years. The main reason for this increased use of longer wavelengths has been to utilize the anomalous signal from sulfur, providing a means for the experimental phasing of native proteins. Here, another possible application of longer X-ray wavelengths is presented: MAD at the L III edges of various lanthanide compounds. A first experiment at the L III edge of Pr was conducted on HZB MX beamline BL14.2 and resulted in the successful structure determination of the C-terminal domain of a spliceosomal protein. This experiment demonstrates that L III edges of lanthanides constitute potentially attractive targets for long-wavelength MAD experiments.

A composite double-/single-stranded RNA-binding region in protein Prp3 supports tri-snRNP stability and splicing

“…Data were processed with the HKL package ( Minor et al, 2006 ) or XDS ( Kabsch, 2010 ). The structure of yPrp3 CTF was solved by a praseodymium (III) multiple anomalous dispersion experiment as described ( Puehringer et al, 2012 ) and refined against the higher resolution data from an yttrium (III)-derivatized crystal. The structures of yPrp3 DUF1115 and of a yPrp3 CTF -yU4/U6 stem II+10nt complex were solved by molecular replacement using structure coordinates of yPrp3 CTF as search models with the programs MOLREP ( Vagin and Teplyakov, 2010 ) and PHASER ( McCoy, 2007 ), respectively.…”

Exploration of the supramolecular interactions involving tris-dipicolinate lanthanide complexes in protein crystals by a combined biostructural, computational and NMR study

“…36 But it has appeared recently in the literature that lanthanide ions are probably among the most promising elements due to their large anomalous signal, which allows phasing a protein of molecular weight 5 times larger. 37,38 Unfortunately, the large majority of proteins are not able to coordinate spontaneously f-block elements. The use of a lanthanide coordination complex rather than a simple salt 39,40 is an interesting strategy, which has first relied on a covalent linkage of the lanthanide recognition tag to the protein backbone.…”