Proteins crystallisation

T.M. Bergfors (Ed.), Protein Crystallisation, International University Line, 1999. ISBN 0963681753. 

Oxford University Press, ISBN 0199636788 (paperback) T.L.Blundell and L.N.Johnson Protein Crystallisation, Academic Press, NY, 1976 A,McPherson Preparation and Analysis of Protein Crystals, J.Wiley Sons, NY, 1982 A.McPherson, Crystallisation of Biological Macromolecules, Cold Spring Harbour Laboratory Press, 2001 ISBN 0879696176.]... 

Chayen, N. E. (2004). Turning protein crystallisation from an art into a science. Curr. Opinion Structural Biol 14, 577-583. 

Chayen, N. E. (2005). Methods for separating nucleation and growth in protein crystallisation. Prog. Biophys. Molec. Biol 88,329-337. 

Garcia-Rufz, J. M., Gonzalez-Ramrrez, L. A., Gavrra, J. A. and OtMora, F. (2002). Granada Crystallisation Box a new device for protein crystallisation by counterdiffusion techniques. Acta Crystallogr. D 58,1638-1642. 

George, A., and Wilson W.W. (1994). Predicting protein crystallisation forma dilute solution property, Cryst. D 50, 361-365. 

Stevens, R.C., High-throughput protein crystallisation. Curr. Opin. Struct. Biol. 2000, 10, 558-563. 

Watanabe, N., Semi-automatic protein crystallisation system that allows in situ observation of X-ray diffraction from crystals in the drop. Acta CrystaUogr. 2002, D58, 1527-1530. 

As a consequence of the individual order of the amino acids and the conformation of the ensuing polypeptide strands (the primary and secondary structures) the three-dimensional structure of each molecule (its tertiary structure) is formed. The bulk of this Chapter is focussed on this aspect of molecular structure. Some comments on quaternary structure and protein crystallisation form a shorter afterword. 

In this context, some comments on protein crystallisation can be made. The process of crystallisation can be viewed as one of self-assembly of the quaternary structure, although the constituent units now have a well-defined arrangement in space, in contrast to their less rigid shape in liquid crystalline mesophases. Indeed, twisted structures are very commonly found in globular protein crystals, which are reminiscent of the hyperbolic forms of micro- and mesoporous zeolites, described in Chapter 2. 

The similar dimensions of many globular proteins and novel mesoporous zeolites offer fascinating possibilities for bio-organic-inorganic composites. For example, the inorganic mesoporous alumino-silicates have been proposed as templates for protein crystallisation [12], Conversely, proteins could be used to template mesostructured inorganic materials. 

Crystallisation was used for many years as a step in the isolation and purification of proteins. Microcrystallinity (the appearance of a silky sheen) was one of the first indications of approaching purity. A good early review of protein crystallisation stemming from this tradition is given by Czok and Bucher [29] for the enzymes of rabbit skeletal muscle. In the past, proteins selected for study were those that were readily available and crystallisable. It is noteworthy that a large number of the enzymes illustrated in Dixon and Webb s [30] crystal atlas have now had their structure solved by X-ray diffraction. 

Poly(ethylene glycol) (PEG) McPherson [35] has advocated the use of PEG for protein crystallisation. Out of 22 proteins chosen to test PEG as a precipitant, 13 yielded crystals, six for the first time. The general formula of PEG is... 

Comparative studies on the same protein crystallised under different conditions and in different space groups show that the structures are essentially the same. For example, monoclinic (C2) subtilisin crystallised from 2.1 M ammonium sulphate, pH 5.9, has the same crystal structure as monoclinic (P2 J subtilisin crystallised from a 55% acetone/water mixture at pH 9.1 [152]. Other examples include tetragonal and triclinic lysozyme, orthorhombic and trigonal trypsin, and trigonal and monoclinic ribonuclease. 

Muller U, Nyarsik L, Horn M, et al (2001) Development of a technology for automation and miniaturisation of protein crystallisation. J Biotechnol (in press)... 

For a satisfactory crystallographic study, it is vital to start with a high quality crystal. This is not easy for proteins. Crystallisation of such large molecular complexes frequently leads to trapped solvent, and a reasonable protein crystal may still contain between 30-80% solvent. Because the... 

For analyte removal, the sensor surface is flushed with an SDS solution, which is an anionic surfactant. Additionally, imprinted layers proved catalytic for protein crystallisation. One could also regard MIP materials as model system of biomineralisation processes D Souza et al. [76] showed an application where crystal-imprinted materials indeed fundamentally increase calcite growth on the surface and therefore work as seeds. 

There are a large number of variables available in a protein crystallisation trial, namely ... 

Figure 3.14 The two basic types of membrane protein ciystals. Type I stacks of membranes containing two-dimensionally crystalline membrane proteins, which are then ordered in the third dimension. Type II a membrane protein crystallised with detergents bound to its hydrophobic surface. The polar surface part of the membrane proteins is indicated by broken lines. The symbols for liquids and detergents are the same as in figure 3.13. From Deisenhofer and Michel (1989) with the permission of the authors, EMBO J, Oxford University Press and copyright The Nobel Foundation (1989).

Crystallisation. The ultimate in purification of proteins or nucleic acids is crystallisation. This involves very specialised procedures and techniques and is best left to the experts in the field of X-ray crystallography who can provide a complete picture of the structure of these large molecules. [A. Ducruix and R. Giege Eds, Crystallisation of Nucleic Acids and Proteins A Practical Approach, 2nd Edition, 2000, Oxford University Press, ISBN 0199636788 (paperback) T.L. Blundell and L.N. Johnson Protein Crystallisation, Academic Press, NY, 197, A. McPherson Preparation and Analysis of Protein Crystals, J.Wiley Sons, NY, 1982, A. McPherson, Crystallisation of Biological Macromolecules, Cold Spring Harbour Laboratory Press, 2001 ISBN 0879696176, see also Bibliography in Chapter 1.]... 

THE UNIQUE POSITION OF THE CORPUSCULAR PROTEINS CRYSTALLISATION AND DENATURATION... 

Now it was already known that many protein crystals contain the micro-units of the mother liquor as well as protein, for example (NH4)2S04 and H2O if the protein crystallised from a sol containing (NH4)2.S04... 

Labbe P Berlot I Lacerda SMV Santos MSCS Lawlor A, McCullagh GD, Zaccarelli E, Eoffi G, Dawson KA Interactions in systems with short-range attractions and applications to protein crystallisation 104... 

Glassford, S.E. et al (2012) Micro ATR FTIR imaging of hanging drop protein crystallisation. Vib, Spectrosc., 63, 492 -498. 

The three-dimensional structure of the PMT active site is not known. The protein structure of a Thermotoga maritima spermidine synthase obtained by protein crystallisation (229) suggested that those amino acids of the SPDS active site that were continuously different between PMTs and SPDS were responsible for the differences in co-substrate acceptance. Mutagenesis of those two amino acids in the D. stramonium PMT with the idea of changing PMT activity into spermidine synthase, however, resulted in a complete loss of catalytic activity (88). A refined protein model of PMT constructed as a consequence of the result discouraged the concept of comparable substrate and co-substrate binding in PMT and SPDS. Rather, the substrate putrescine binds differentially at the active sites of both enzymes. 

Ion specific effects generally follow direct or reverse order of the so-called Hofineister series, which for monovalent anions is, SCN > CIO4 1 > Br- > Cl- > F. In many cases, interfacial effects appear to play a key role. In the simplest case of ions at the air-water interface for example, the specificity is directly reflected in the stability of foams or in the surface tension and surface potential of aqueous salt solutions. In biology, beyond protein crystallisation, the enzymatic activity can be controlled by addition of alkali halides with large differences between fluoride, chloride, bromide, or iodide. Specific types of cations are also identified together with... 

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