Crystallization, X-ray diffraction analysis and SIRAS phasing of human ?-L-iduronidase.

ABSTRACT: Human lysosomal ?-L-iduronidase, whose deficiency causes mucopolysaccharidosis type I, was crystallized using sodium/potassium tartrate and polyethylene glycol 3350 as a precipitant. Using synchrotron radiation, a native data set was collected from a single crystal at 100?K to 2.3?Å resolution. The crystal belonged to space group R3 with unit-cell dimensions of a=b=259.22, c=71.83?Å. To obtain the phase information, mercury-derivative crystals were prepared and a single-wavelength anomalous dispersion (SAD) data set was collected at the Hg peak wavelength. Phase calculation with the single isomorphous replacement with anomalous scattering (SIRAS) method successfully yielded an interpretable electron-density map.

SUBMITTER: Maita N 

PROVIDER: S-EPMC3515382 | biostudies-literature | 2012 Nov

REPOSITORIES: biostudies-literature