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Preparing recombinant proteins for X-ray crystallography
Hughes, S. H. and Stock, A. M.  International Tables for Crystallography (2012). Vol. F, ch. 3.1, pp. 75-91 [ doi:10.1107/97809553602060000810 ]
... extant problems and the available solutions, so that, armed with a general understanding of the issues, they can more easily confront a variety of specific projects. Specific topics covered include protein engineering ... Preparing protein crystals appropriate for X-ray diffraction usually requires a considerable amount of highly purified protein. When crystallographic methods ...

Protein storage
Hughes, S. H. and Stock, A. M.  International Tables for Crystallography (2012). Vol. F, Section 3.1.6.2, p. 89 [ doi:10.1107/97809553602060000810 ]
... under the best of circumstances, protein solutions are subject to a number of unwanted events that can include, but are not ... limited to, oxidation, racemization, deamination, denaturation, proteolysis and aggregation. As a general rule, it is better to store proteins as highly ... crystallization. If the protein contains oxidizable sulfurs (free cysteines are a particular problem), reducing agents can be added (and should ...
     [more results from section 3.1.6 in volume F]

Purifying and refolding denatured proteins
Hughes, S. H. and Stock, A. M.  International Tables for Crystallography (2012). Vol. F, Section 3.1.5.3, pp. 87-88 [ doi:10.1107/97809553602060000810 ]
... not with the purification of the protein, but in finding a proper way to refold it. Various general procedures for refolding ... specific protocols. The insoluble inclusion bodies are usually solubilized in a powerful chaotropic agent like guanidine hydrochloride or urea. In general ... in the buffer to accelerate correct pairing of disulfides. After a refolding procedure, the properly folded soluble protein must be ...
     [more results from section 3.1.5 in volume F]

Mammalian cells
Hughes, S. H. and Stock, A. M.  International Tables for Crystallography (2012). Vol. F, Section 3.1.4.4, pp. 84-85 [ doi:10.1107/97809553602060000810 ]
... prepared by transfection; following transfection, some of the cells (usually a small percentage) will incorporate transfected DNA into their genomes. A number of agents can be used to transfect DNA; these ... limited to, CaPO4, DEAE Dextran, cationic lipids etc. This is a complex and poorly defined process; the transfected DNA is ...
     [more results from section 3.1.4 in volume F]

Addition of tags or domains
Hughes, S. H. and Stock, A. M.  International Tables for Crystallography (2012). Vol. F, Section 3.1.3.3, p. 77 [ doi:10.1107/97809553602060000810 ]
... or domains In some cases it is useful to add a small peptide tag or a larger protein to either the amino or carboxyl terminus of ... tags, such as the maltose-binding protein, thioridazine, and protein A, can also act as molecular chaperones to aid in ...
     [more results from section 3.1.3 in volume F]

Overview
Hughes, S. H. and Stock, A. M.  International Tables for Crystallography (2012). Vol. F, Section 3.1.2, pp. 75-76 [ doi:10.1107/97809553602060000810 ]
... idea that underlies the problem of expressing large amounts of a recombinant protein is straightforward: prepare a DNA segment that, when introduced into an appropriate host, will ... other ways that are relevant to crystallography. In some cases, a protein in its natural form is not suitable for ...

Introduction
Hughes, S. H. and Stock, A. M.  International Tables for Crystallography (2012). Vol. F, Section 3.1.1, p. 75 [ doi:10.1107/97809553602060000810 ]
... Preparing protein crystals appropriate for X-ray diffraction usually requires a considerable amount of highly purified protein. When crystallographic methods were ... techniques changed the rules: it is now possible to instruct a variety of cells and organisms to make large amounts of ... not our intention in writing this chapter to provide either a methods manual for those interested in expressing a particular ...

Reprise
Hughes, S. H. and Stock, A. M.  International Tables for Crystallography (2012). Vol. F, Section 3.1.7, pp. 89-90 [ doi:10.1107/97809553602060000810 ]
Reprise 3.1.7. Reprise We have reached a point where it is possible to use recombinant DNA techniques ... proteins vastly simplify the process of purification. This has played a direct and critical role in the ability of crystallographers to ... sequence of proteins to improve their crystallization properties. This is a difficult problem; however, there are already notable, if hard ...

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