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PDBsum entry 2evz
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RNA binding protein
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PDB id
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2evz
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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 the two most c-Terminal RNA recognition motifs of ptb using segmental isotope labeling.
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Authors
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F.Vitali,
A.Henning,
F.C.Oberstrass,
Y.Hargous,
S.D.Auweter,
M.Erat,
F.H.Allain.
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Ref.
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EMBO J, 2006,
25,
150-162.
[DOI no: ]
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PubMed id
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Abstract
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The polypyrimidine tract binding protein (PTB) is a 58 kDa protein involved in
many aspects of RNA metabolism. In this study, we focused our attention on the
structure of the two C-terminal RNA recognition motifs (RRM3 and RRM4) of PTB.
In a previous study, it was found that the two RRMs are independent in the free
state. We recently determined the structure of the same fragment in complex with
RNA and found that the two RRMs interact extensively. This difference made us
re-evaluate in detail the free protein structure and in particular the
interdomain interface. We used a combination of NMR spectroscopy and segmental
isotopic labeling to unambiguously study and characterize the interdomain
interactions. An improved segmental isotopic labeling protocol was used,
enabling us to unambiguously identify 130 interdomain NOEs between the two RRMs
and to calculate a very precise structure. The structure reveals a large
interdomain interface, resulting in a very unusual positioning of the two RRM
domains relative to one another.
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Figure 1.
Figure 1 Protein constructs used in this study. Protein domains
are represented by colored boxes: orange for the NLS sequence;
white for linker regions; pink for PTB RRM1; violet for PTB
RRM2; red for PTB RRM3 domain; yellow for PTB RRM4; gray for 6
 His
tag; blue for CBD; light blue for the Mxe GyrA intein; and green
for the Ssp DnaB intein.
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Figure 6.
Figure 6 Close-up view of the interdomain interface in PTB RRM34
and hnRNPA1 RRM12. (A) Stereo view of the interaction between
helix 2 of RRM4 and helix 1 of RRM3. Side chains for RRM3 and
RRM4 and the interdomain linker are represented by sticks and
dotted surfaces colored in blue, green and red, respectively.
(B) Stereo view of the interaction between helix 2 of RRM3 with
the interdomain linker and F526 from RRM4. (C) Stereo view of
the interdomain interface found in the structure of hnRNPA1
RRM12 (Shamoo et al, 1997).
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The above figures are
reprinted
by permission from Macmillan Publishers Ltd:
EMBO J
(2006,
25,
150-162)
copyright 2006.
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