The science of macromolecular crystallization

Abstract

For the X-ray crystallographer, the future of his field is, and has always been, determined by what he is able to crystallize. This is now particularly true as a result of the extraordinary improvement in X-ray detectors and sources [1], the advent of computers of amazing speed, and the development of programs that are both friendly and efficient [2]. The critical component, trailing other technology, is the growth of crystals of macromolecules having sufficient size and quality to permit X-ray analysis.Crystallization, however, has become less problematic. With synchrotron sources [3], sensitive, fast detectors, cryogenic techniques that eliminate radiation damage [4], and more powerful phasing tools, both the number and the size of crystals required for the analysis has decreased: crystals of 30-50 ,pm may soon be adequate, frozen crystals often provide a complete data set, and molecular replacement can allow determination of entire structures from this one set. Equally importantly, crystallization procedures, reagents, and diagnostic tools are now available that greatly improve the probability of success for both experienced crystallographers and interested biochemists.

Introduction to protein crystallization

“…There are further differences which complicate the crystallization of macromolecules compared with conventional small molecules (Feigelson, 1988;Feher, 1986;Durbin & Feher, 1996;McPherson, 1982McPherson, , 1999McPherson et al, 1995). Firstly, macromolecules may assume several distinctive solid states that include amorphous precipitates, oils or gels as well as crystals, and most of these are kinetically favored.…”

Applications of the second virial coefficient: protein crystallization and solubility

“…However, in addition to microgravity studies, each space agency also provided funding to support ground-based studies addressing the fundamental aspects of protein crystal growth. Some of these studies are directed at understanding the causes of different crystal defects, crystal growth termination, how crystal growth rate affects crystal quality, dynamic control of the crystal nucleation and growth phase, and investigations of how fluid flows and protein transport rate influence crystal size and quality (Li et al, 1999a,b;Forsythe et al, 2002;Gorti et al, 2005;Vekilov, 2003Vekilov, , 2009Vekilov, , 2010Booth et al, 2004;Thomas et al, 1996Thomas et al, , 1998McPherson et al, 1995McPherson et al, , 2001Malkin & McPherson, 1994;Kuznetsov et al, 1997Kuznetsov et al, , 2000Drenth & Haas, 1998;Vekilov et al, 1996). Other advances in protein crystallization strategy include the statistical design of experiments, analysis of screen results, automated crystal image analysis and novel seeding approaches Luft, Wolfley et al, 2011;Nagel et al, 2008;Snell et al, 2008;D'Arcy et al, 2003D'Arcy et al, , 2004D'Arcy et al, , 2007.…”

A guide to the crystallographic analysis of icosahedral viruses

“…Face normal growth seems to be exclusively due to two-and three-dimensional nucleation on existing surfaces. [58,98] Virus crystals, because of the large growth step heights at the advancing edges ( Figure 10), incorporate vast amounts of impurities into their lattices. That is, the growth steps, as they move across the surfaces of crystals, singly or in step bunches, [99] sweep everything before them, like great waves, into the channels and interstices between particles.…”