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PDBsum entry 1m8m
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Structural protein
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PDB id
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1m8m
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Contents |
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* Residue conservation analysis
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References listed in PDB file
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Key reference
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Title
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Structure of a protein determined by solid-State magic-Angle-Spinning nmr spectroscopy.
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Authors
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F.Castellani,
B.Van rossum,
A.Diehl,
M.Schubert,
K.Rehbein,
H.Oschkinat.
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Ref.
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Nature, 2002,
420,
98.
[DOI no: ]
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PubMed id
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Abstract
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The determination of a representative set of protein structures is a chief aim
in structural genomics. Solid-state NMR may have a crucial role in structural
investigations of those proteins that do not easily form crystals or are not
accessible to solution NMR, such as amyloid systems or membrane proteins. Here
we present a protein structure determined by solid-state magic-angle-spinning
(MAS) NMR. Almost complete (13)C and (15)N resonance assignments for a
micro-crystalline preparation of the alpha-spectrin Src-homology 3 (SH3) domain
formed the basis for the extraction of a set of distance restraints. These
restraints were derived from proton-driven spin diffusion (PDSD) spectra of
biosynthetically site-directed, labelled samples obtained from bacteria grown
using [1,3-(13)C]glycerol or [2-(13)C]glycerol as carbon sources. This allowed
the observation of long-range distance correlations up to approximately 7 A. The
calculated global fold of the alpha-spectrin SH3 domain is based on 286
inter-residue (13)C-(13)C and six (15)N-(15)N restraints, all self-consistently
obtained by solid-state MAS NMR. This MAS NMR procedure should be widely
applicable to small membrane proteins that can be expressed in bacteria.
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Figure 2.
Figure 2: Assignment strategy. Regions extracted from the
spectra of Fig. 1c and d (I -VI) superimposed on a PDSD spectrum
of uniformly labelled SH3 domain (black), the latter recorded
with a short mixing time of 15 ms (ref. 5. Part of the
assignment of the long-range correlations is reported in the
figure, and the lines define the different correlation patterns.
As an example, correlations between residues L33 and V44 are
shown. The correlations between the C  and
C  signals
of V44 and C and
C  signals
of L33 are observed in panels I -III for 2-SH3, whereas
correlations between the methyl groups appear in the spectrum of
1,3-SH3 (panel IV). Of particular interest is the region around
50 p.p.m. (panel VI), where for 2-SH3 a large number of
cross-peaks due to the proline-  signals
(P20 and P54) are observed, whereas in the corresponding area of
the U-SH3 sample (see Fig. 1b), no correlations are detected. In
the upper left corner, a schematic representation of an
antiparallel -sheet
is shown. The numbering of the residues (i and j) corresponds to
the numbering in Table 1.
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Figure 3.
Figure 3: Solid-state structure of the alpha- -spectrin
SH3 domain. a, Stereo view of twelve of the fifteen
lowest-energy structures, representing the fold of the SH3
domain. The three structures with the largest r.m.s. deviation
to the average structure are not displayed. The -strand
regions are shown in blue. b, The X-ray structure^26 is shown
for comparison. In this case, the part of the -sheet
in the region 14 -17 and 23 -26 is non-ideal and therefore is
not indicated in blue.
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The above figures are
reprinted
by permission from Macmillan Publishers Ltd:
Nature
(2002,
420,
98-0)
copyright 2002.
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