Purification and crystallization of the ternary complex of elongation factor Tu:GTP and Phe-tRNA(Phe).
Elongation factor Tu (EF-Tu) is the most abundant protein in prokaryotic cells. Its general function in protein biosynthesis is well established. It is a member of the large family of G-proteins, all of which bind guanosine phosphates (GDP or GTP) as cofactors. In its active GTP bound state EF-Tu binds aminoacylated tRNA (aa-tRNA) forming the ternary complex EF-Tu:GTP:aa-tRNA. The ternary complex interacts with the ribosome where the anticodon on tRNA recognises a codon on mRNA, GTPase activity is induced and inactive EF-Tu:GDP is released. Here we report the successful crystallization of a ternary complex of Thermus aquaticus EF-Tu:GDPNP and yeast Phe-tRNA(Phe) after its purification by HPLC.
Nissen
P
P
Department of Biostructural Chemistry, University of Aarhus, Denmark.
Reshetnikova
L
L
Siboska
G
G
Polekhina
G
G
Thirup
S
S
Kjeldgaard
M
M
Clark
B F
BF
Nyborg
J
J
D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D003460:Crystallization; D018360:Crystallography, X-Ray; D004591:Electrophoresis, Polyacrylamide Gel; D006160:Guanosine Triphosphate / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D006165:Guanylyl Imidodiphosphate / Q000378:metabolism; D010444:Peptide Elongation Factor Tu / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D012360:RNA, Transfer, Phe / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D013824:Thermus / Q000378:metabolism
D012360:RNA, Transfer, Phe; D006165:Guanylyl Imidodiphosphate; D006160:Guanosine Triphosphate; D010444:Peptide Elongation Factor Tu