Significance: The observation of dynamical diffraction in protein crystals is an interesting topic because dynamical diffraction typically occurs in high-quality crystals such as those of silicon. However, to our knowledge, a report of protein crystals showing clear dynamical diffraction does not exist. Here, we present an observation of the oscillatory profile of rocking curves for protein crystals such as glucose isomerase. The oscillatory profiles are in good agreement with those predicted using the dynamical theory of diffraction. It is demonstrated that dynamical diffraction occurs in protein crystals as well. These results indicate the need for a dynamical diffraction model in protein-crystal structure analysis and model refinement that, to our knowledge, has never been used in conventional structural analysis.

Hardness and slip systems by an indentation method were investigated on different habit planes of orthorhombic hen egg-white lysozyme (O-HEWL) crystals containing water. A dependence of the hardness on the water-evaporation time exhibits three stages as incubation, transition and saturated ones, as tetragonal (T)-HEWL crystals reported previously. The hardness values of (1 1 0), (0 1 0) and (0 1 1) habit planes of O-HEWL in the incubation stage or wet condition exhibits 6, 8 and 10 MPa, respectively. The hardness depends on indented planes but it is independent of the air-humidity and crystal volumes. These values correspond to the intrinsic hardness for O-HEWL crystals containing water. In the incubation stage, the slip traces are clearly observed around the indentation mark and the corresponding six kinds of slip systems are identified to be {0 1 1}, {1 1 0}, {0 1 1}, {1 1 0}, {1 0 0} and {0 1 0}.

Knowledge of X-ray diffraction in macromolecular crystals is important for not only structural analysis of proteins but also diffraction physics. Dynamical diffraction provides evidence of perfect crystals. Until now, clear dynamical diffraction in protein crystals has only been observed in glucose isomerase crystals. We wondered whether there were other protein crystals with high quality that exhibit dynamical diffraction. Here we report the observation of dynamical diffraction in thin ferritin crystals by rocking-curve measurement and imaging techniques such as X-ray topography. It is generally known that in the case of thin crystals it is difficult to distinguish whether dynamical diffraction occurs from only rocking-curve profiles. Therefore, our results clarified that dynamical diffraction occurs in thin protein crystals because fringe contrasts similar to Pendellösung fringes were clearly observed in the X-ray topographic images. For macromolecular crystallography, it is hard to obtain large crystals because they are difficult to crystallize. For thin crystals, dynamical diffraction can be demonstrated by analysis of the equal-thickness fringes observed by X-ray topography.

A long alkyl-chained organic molecule, 4,5,6,7-tetrachloro-2-dodecylisoindoline-1,3-dione (1), was crystallized into needle-like crystals in dichloromethane (1^DCM) or plate-like ones in tetrahydrofuran (1^THF) depending on the recrystallisation solvent. X-ray crystallography analyses revealed the alkyl chains of the molecules, in which they were assembled differently, with the former responding flexibly bendable and elastic deformation, and the later being a permanent plastic one by external mechanical stress. The elastic modulus (E) and hardness (H) indicating both compliant and soft nature, reflecting their weak interaction in crystals, were quantified from the nano-indentation test.