Progress in protein crystallography

Abstract

Macromolecular crystallography evolved enormously from the pioneering days, when structures were solved by “wizards” performing all complicated procedures almost by hand. In the current situation crystal structures of large systems can be often solved very effectively by various powerful automatic programs in days or hours, or even minutes. Such progress is to a large extent coupled to the advances in many other fields, such as genetic engineering, computer technology, availability of synchrotron beam lines and many other techniques, creating the highly interdisciplinary science of macromolecular crystallography. Due to this unprecedented success crystallography is often treated as one of the analytical methods and practiced by researchers interested in structures of macromolecules, but not highly competent in the procedures involved in the process of structure determination. One should therefore take into account that the contemporary, highly automatic systems can produce results almost without human intervention, but the resulting structures must be carefully checked and validated before their release into the public domain.

Mesoscale imaging with cryo‐light and X‐rays: Larger than molecular machines, smaller than a cell

“…An army of researchers have applied techniques such as electron microscopy (EM) (Nogales ), nuclear magnetic resonance (NMR) (Dutta et al. ) and macromolecular X‐ray crystallography (Dauter and Wlodawer, ) to determine the tertiary structures of nucleotides, proteins, and even viruses. The effort in this area is on‐going, and rapidly expanding in the case of cryo‐EM, where a resurgence in the technique has a focus on determining the structure of ever more complex molecules, including multi‐component molecular machines at near‐atomic resolution (Nogales ).…”

Paradoxical effects on voltage-gated Na+ conductance in adrenal chromaffin cells by twin vs single high intensity nanosecond electric pulses

“…X-ray diffraction imaging, and the development of new computer algorithms to reconstruct images [47,48]. Despite these developments, our understanding of the properties of transmembrane proteins, and in particular ion channels in their native environment, still remains limited because the vast majority of protein structures that have been uncovered were determined without the stabilizing effects of a surrounding lipid bilayer.…”

Protein X-ray Crystallography and Drug Discovery

“…The core of this cycle still applies nowadays, although with more sophistication (Figure 1). With the advent of crystallization automates, brighter synchrotron X-ray sources, faster detectors, automated structure solution, and refinement pipelines, it is not uncommon to be able to determine macromolecular structures within a few days [10], and in the case of structures of protein-ligand complexes, throughput of several structures a day is attainable [11]. As the throughput augmented, the use of X-ray crystallography progressed from the protein target structure determination, possibly in the presence of some pre-identified ligand, to structure-activity relationship determination by X-ray crystallography, where several structures of complexes are determined in order to guide ligand optimization [12], to a screening tool of chemical libraries containing a few hundreds of compounds distributed as cocktails of up to ten compounds [13] or even individually as it is now feasible [14].…”