Solution structure of the ternary complex between aminoacyl-tRNA, elongation factor Tu, and guanosine triphosphate.
Complex formation between elongation factor Tu (EF-Tu), Phe-tRNAPhe, and GTP was analyzed by small-angle neutron and X-ray scattering methods. Both techniques show that the ternary complex consists of one EF-Tu and one aminoacyl-tRNA. No shift in stoichiometry was detected when the temperature was raised from 5 to 37 degreesC, in contrast to previous observations obtained from RNase A protection experiments [Bilgin and Ehrenberg (1995) Biochemistry34, 715-719]. A small but significant increase in the radius of gyration of the complex was observed when the temperature was decreased from 37 to 5 degreesC. The X-ray solution scattering patterns were compared with those calculated from the crystal structure of the complex formed between EF-Tu from Thermus aquaticus and Phe-tRNAPhe from yeast. The comparison shows that the solution structure of the ternary complex, formed entirely from Escherichia coli components and under translationally optimal buffer conditions, is very close to the crystal structure, formed from heterologous components under very different conditions. Furthermore, for the hybrid complex in solution there is no evidence for the formation of trimers as suggested by the crystal structure.
Bilgin
N
N
Department of Molecular Biology, The Biomedical Center, Uppsala University, Sweden. bilgin@xray.bmc.uu.se
Ehrenberg
M
M
Ebel
C
C
Zaccai
G
G
Sayers
Z
Z
Koch
M H
MH
Svergun
D I
DI
Barberato
C
C
Volkov
V
V
Nissen
P
P
Nyborg
J
J
D001665:Binding Sites; D018360:Crystallography, X-Ray; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000737:chemistry; D006160:Guanosine Triphosphate / Q000737:chemistry*; D046911:Macromolecular Substances; D008958:Models, Molecular; D009502:Neutrons; D010444:Peptide Elongation Factor Tu / Q000737:chemistry* / Q000378:metabolism; D012360:RNA, Transfer, Phe / Q000737:chemistry* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000737:chemistry; D012542:Scattering, Radiation; D012996:Solutions; D017356:Synchrotrons; D013824:Thermus / Q000737:chemistry; D014965:X-Rays
D046911:Macromolecular Substances; D012360:RNA, Transfer, Phe; D012996:Solutions; D006160:Guanosine Triphosphate; D010444:Peptide Elongation Factor Tu