added a model of C73-C74 of the tRNA and residues 342–351 and 375–390 of Aa.LC. The
refinement was carried out by conjugate gradient minimization and torsion angle
dynamics simulated annealing
30
. Iterative model building and refinement improved the
model, resulting in final R
work
and R
free
values of 23.0% and 28.7%, respectively.
The final model includes two AMPcPP molecules and 79 water molecules. All of the
refinement calculations were done with CNS
31
. Refinement statistics are shown in
Supplementary Table 1. All of the figures were prepared with the programs Que
(http://www.biochem.s.u-tokyo.ac.jp/,ishitani/que/) and PyMol (http://
pymol.sourceforge.net/).
In vitro assays
Mutations were generated by the QuickChange mutagenesis kit (Stratagene). The mutant
proteins were overexpressed and purified as described for the wild-type enzyme. We
assayed activity in a solution containing 50 mM glycine (pH 8.5), 10 mM MgCl
2
, 35 mM
KCl, 1 mM dithiotheitol, 5% (v/v) glycerol, 0.2 mM tRNA, 10 mM ATP, 100 nM [a-
32
P]ATP
(3,000 Ci mol
21
) and 10 nM enzyme at 50 8C. After the reactions were stopped, the
samples were separated by 10% denaturing PAGE. The assay conditions were developed,
on the basis of the wild-type Michaelis constant (K
m
) values for tRNA (2.3 mM) and ATP
(47 mM), to be sensitive enough to evaluate mutant activity. Assays of mutant tRNAs
(tRNA-N74N75, where N is A, G, C or U) were done as described above, except that the
concentrations of tRNA-N74N75 and ATP were 5 and 250 mM, respectively.
Received 6 April; accepted 4 June 2004; doi:10.1038/nature02712.
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Supplementary Information accompanies the paper on www.nature.com/nature.
Acknowledgements We thank A. M. Weiner for the Aa.LC plasmids; and M. Kawamoto and
H. Sakai for help with data collection at SPring-8. This work was supported by Kurata Memorial
Hitachi Science and Technology Foundation, Takeda Science Foundation, Foundation of
Advanced Technology Institute and a Grant-in-aid for Young Scientists (to K.T.); by Asahi Glass
Foundation (to S.F.); by a grant from the Ministry of Education, Culture, Sports, Science and
Technology (to N.T.); and by a PRESTO Program grant from Japan Science and Technology and a
Naito Foundation grant (to O.N.).
Competing interests statement The authors declare that they have no competing financial
interests.
Correspondence and requests for materials should be addressed to O.N.
(onureki@bio.titech.ac.jp). Atomic coordinates have been deposited in the Protein Data Bank
under accession number 1VFG.
..............................................................
erratum
Single-crystal metallic nanowires
and metal/semiconductor nanowire
heterostructures
Yue Wu, Jie Xiang, Chen Yang, Wei Lu & Charles M. Lieber
Nature 430, 61–65 (2004).
.............................................................................................................................................................................
In Fig. 2b of this Letter, the units of the abscissa should be volts (V).
In Fig. 3a, the step labels (3) and (4) should be interchanged. A
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