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PDBsum entry 1x02
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Metal binding protein
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PDB id
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1x02
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References listed in PDB file
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Key reference
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Title
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Optimal isotope labelling for nmr protein structure determinations.
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Authors
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M.Kainosho,
T.Torizawa,
Y.Iwashita,
T.Terauchi,
A.Mei ono,
P.Güntert.
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Ref.
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Nature, 2006,
440,
52-57.
[DOI no: ]
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PubMed id
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Abstract
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Nuclear-magnetic-resonance spectroscopy can determine the three-dimensional
structure of proteins in solution. However, its potential has been limited by
the difficulty of interpreting NMR spectra in the presence of broadened and
overlapping resonance lines and low signal-to-noise ratios. Here we present
stereo-array isotope labelling (SAIL), a technique that can overcome many of
these problems by applying a complete stereospecific and regiospecific pattern
of stable isotopes that is optimal with regard to the quality and information
content of the resulting NMR spectra. SAIL uses exclusively chemically and
enzymatically synthesized amino acids for cell-free protein expression. We
demonstrate for the 17-kDa protein calmodulin and the 41-kDa
maltodextrin-binding protein that SAIL offers sharpened lines, spectral
simplification without loss of information, and the ability to rapidly collect
the structural restraints required to solve a high-quality solution structure
for proteins twice as large as commonly solved by NMR. It thus makes a large
class of proteins newly accessible to detailed solution structure determination.
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Figure 1.
Figure 1: SAIL amino acids. Design concepts embodied in the
SAIL amino acids incorporated into CaM and MBP^13–15.
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Figure 4.
Figure 4: CaM and MBP solution and crystal structures.
Figure 4 : CaM and MBP solution and crystal structures.
Unfortunately we are unable to provide accessible
alternative text for this. If you require assistance to access
this image, or to obtain a text description, please contact
npg@nature.com-
a, SAIL-CaM (backbone in cyan, Ca^2+ in white), CaM X-ray
structure^31 (red), and three solution conformers of UL-CaM
determined from residual dipolar coupling data^32 (blue). b, MBP
solution and crystal structures^27. Backbone of the N-terminal
domain (SAIL-MBP in green, X-ray in red) and the C-terminal
domain (SAIL-MBP in blue, X-ray in red). c, Aromatic side chains
(SAIL-MBP in green and blue, X-ray in red), and backbone ribbon
representation of the X-ray structure (gold). Superpositions of
CaM and MBP solution conformers on the X-ray structures were
performed separately for the two flexibly connected domains.
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The above figures are
reprinted
by permission from Macmillan Publishers Ltd:
Nature
(2006,
440,
52-57)
copyright 2006.
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