Open AccessCrystallization of Macromolecules
Author(s) -
Friedmann David,
Messick Troy,
Marmorstein Ronen
Publication year - 2011
Publication title -
current protocols in protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.409
H-Index - 32
eISSN - 1934-3663
pISSN - 1934-3655
DOI - 10.1002/0471140864.ps1704s66
Subject(s) - macromolecule , crystallization , nucleic acid , macromolecular substances , protein crystallization , reagent , chemistry , crystallography , materials science , biochemistry , organic chemistry
X‐ray crystallography has evolved into a very powerful tool to determine the three‐dimensional structure of macromolecules and macromolecular complexes. The major bottleneck in structure determination by X‐ray crystallography is the preparation of suitable crystalline samples. This unit outlines steps for the crystallization of a macromolecule, starting with a purified, homogeneous sample. The first protocols describe preparation of the macromolecular sample (i.e., proteins, nucleic acids, and macromolecular complexes). The preparation and assessment of crystallization trials is then described, along with a protocol for confirming whether the crystals obtained are composed of macromolecule as opposed to a crystallization reagent. Next, the optimization of crystallization conditions is presented. Finally, protocols that facilitate the growth of larger crystals through seeding are described. Curr. Protoc. Protein Sci . 66:17.4.1‐17.4.26. © 2011 by John Wiley & Sons, Inc.