Growth and disorder of macromolecular crystals: insights from atomic force microscopy and X-ray diffraction studies

• Published in: Methods (San Diego, Calif.) Vol. 34; no. 3; pp. 273 - 299
• Main Authors: Malkin, Alexander J., Thorne, Robert E.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01-11-2004
• Subjects: Atomic force microscopy; Chemistry Techniques, Analytical - methods; Crystal growth; Crystallization; Crystallography; Cytomegalovirus - chemistry; Defect structure; Disorder; Growth mechanisms; High-resolution reciprocal space scanning; Macromolecular crystals; Microscopy, Atomic Force - instrumentation; Microscopy, Atomic Force - methods; Plant Proteins - chemistry; Protein crystals; Proteins - chemistry; Structural genomics; Temperature; Two-dimensional nucleation; Virus crystals; Viruses - chemistry; X-Ray Diffraction - methods; X-ray diffraction topography; Crystal growth; Atomic force microscopy; Growth mechanisms; Macromolecular crystals; Protein crystals; Virus crystals; Structural genomics; Crystallography; Disorder; Two-dimensional nucleation; High-resolution reciprocal space scanning; X-ray diffraction topography; Defect structure
• ISSN: 1046-2023; 1095-9130
• DOI: 10.1016/j.ymeth.2004.03.020

The growth processes and defect structures of protein and virus crystals have been studied in situ by atomic force microscopy (AFM), X-ray diffraction topography, and high-resolution reciprocal space scanning. Molecular mechanisms of macromolecular crystallization were visualized and fundamental kinetic and thermodynamic parameters, which govern the crystallization process of a number of macromolecular crystals, have been determined. High-resolution AFM imaging of crystal surfaces provides information on the packing of macromolecules within the unit cell and on the structure of large macromolecular assemblies. X-ray diffraction techniques provide a bulk probe with poorer spatial resolution but excellent sensitivity to mosaicity and strain. Defect structures and disorder created in macromolecular crystals during growth, seeding, and post-growth treatments including flash cooling were characterized and their impacts on the diffraction properties of macromolecular crystals have been analyzed. The diverse and dramatic effects of impurities on growth and defect formation have also been studied. Practical implications of these fundamental insights into the improvement of macromolecular crystallization protocols are discussed.