Journal of Synchrotron Radiation

Volume 15, Part 3 (May 2008)

diffraction structural biology

J. Synchrotron Rad. (2008). 15, 262-265 [ doi:10.1107/S0909049507056531 ]

Crystal twinning of human MD-2 recognizing endotoxin cores of lipopolysaccharide

U. Ohto and Y. Satow

Abstract: Twinning of crystals causes overlapping of two or more reciprocal lattice points, and hence structure amplitudes for a single crystalline domain are hardly obtained from X-ray diffraction intensities. MD-2 protein forms a stable complex with Toll-like receptor 4 and recognizes bacterial lipopolysaccharide (LPS). Excessive immune responses activated by LPS cause septic shocks. Saccharide-trimmed human MD-2 crystallizes in the tetragonal form with apparent Laue symmetry of 4/mmm, and diffraction intensities from these crystals indicate crystal twinning. The crystal consists of two different domains, A and B. The c_A axis of domain A coincides with the c_B axis of domain B with a smaller lattice, and the a_A axis corresponds to the (a_B + b_B) axis. This twinning severely imposes difficulty in structure determination. Through optimization of cryoprotectant, domain A was thoroughly transformed into domain B. The crystal containing only domain B is in space group P4₁2₁2 with one MD-2 molecule in the asymmetric unit. The structure of this form of MD-2 as well as its complex with antiendotoxic lipid IVa was successfully determined using the multiple isomorphous replacement method.

Keywords: crystal twinning; innate immunity; endotoxin.