TY - CHAP
T1 - Crystallization of RNA-protein complexes
T2 - From synthesis and purification of individual components to crystals
AU - Perederina, Anna
AU - Krasilnikov, Andrey S.
PY - 2012
Y1 - 2012
N2 - A broad range of biological processes relies on complexes between RNA and proteins. Crystallization of RNA-protein complexes can yield invaluable information on structural organizations of key elements of cellular machinery. However, crystallization of RNA-protein complexes is often challenging and requires special approaches. Here we review the purification of RNA, RNA-binding proteins, and the formation and crystallization of RNA-protein complexes, using the crystallization of the P3 RNA domain of ribonuclease MRP, a multicomponent ribonucleoprotein complex involved in the metabolism of various RNA molecules, as an example. The RNA-protein complex was formed using gel-purified RNA, produced by run-off transcription with T7 RNA polymerase in vitro, and proteins that were overexpressed in Escherichia coli and purified to be RNase-free. The complex was crystallized using a sitting drop setup; initial screening for suitable crystallization conditions was performed using a sparse matrix approach.
AB - A broad range of biological processes relies on complexes between RNA and proteins. Crystallization of RNA-protein complexes can yield invaluable information on structural organizations of key elements of cellular machinery. However, crystallization of RNA-protein complexes is often challenging and requires special approaches. Here we review the purification of RNA, RNA-binding proteins, and the formation and crystallization of RNA-protein complexes, using the crystallization of the P3 RNA domain of ribonuclease MRP, a multicomponent ribonucleoprotein complex involved in the metabolism of various RNA molecules, as an example. The RNA-protein complex was formed using gel-purified RNA, produced by run-off transcription with T7 RNA polymerase in vitro, and proteins that were overexpressed in Escherichia coli and purified to be RNase-free. The complex was crystallized using a sitting drop setup; initial screening for suitable crystallization conditions was performed using a sparse matrix approach.
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U2 - 10.1007/978-1-61779-949-5_9
DO - 10.1007/978-1-61779-949-5_9
M3 - Chapter
C2 - 22736002
AN - SCOPUS:84864142010
SN - 9781617799488
T3 - Methods in Molecular Biology
SP - 123
EP - 143
BT - Bacterial Regulatory RNA
A2 - Keiler, Kenneth C.
ER -