Quail Cysteine and Glycine-rich Protein, NMR, 15 Minimized Model Structures

1QLI: U-15N isotopical labelling but no U-13C isotopical labelling

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Application of cross-correlated NMR spin relaxation to the zinc-finger protein CRP2(LIM2): evidence for collective motions in LIM domains.

Proteins of the cysteine-rich protein (CRP) family (CRP1, CRP2, and CRP3) are
implicated in diverse processes linked to cellular differentiation and growth
control. CRP proteins contain two LIM domains, each formed by two zinc-binding
modules of the CCHC and CCCC type, respectively. The solution structure of the
carboxyl-terminal LIM domain (LIM2) from recombinant quail CRP2 was determined
by multidimensional homo- and heteronuclear magnetic resonance spectroscopy.
The folding topology retains both independent zinc binding modules (CCHC and
CCCC). Each module consists of two orthogonally arranged antiparallel
beta-sheets, and the carboxyl-terminal CCCC module is terminated by an
alpha-helix. 15N magnetic relaxation data indicate that the modules differ in
terms of conformational flexibility. They pack together via a hydrophobic core
region. In addition, Arg122 in the CCHC module and Glu155 in the CCCC module
are linked by an intermodular hydrogen bond and/or salt bridge. These residues
are absolutely conserved in the CRP family of LIM proteins, and their
interaction might contribute to the relative orientation of the two
zinc-binding modules in CRP LIM2 domains. The global fold of quail CRP2 LIM2 is
very similar to that of the carboxyl-terminal LIM domain of the related but
functionally distinct CRP family member CRP1, analyzed recently. The
carboxyl-terminal CCCC module is structurally related to the DNA-binding domain
of the erythroid transcription factor GATA-1. In the two zinc-binding modules
of quail CRP2 LIM2, flexible loop regions made up of conserved amino acid
residues are located on the same side of the LIM2 domain and may cooperate in
macromolecular recognition.

Konrat R, Weiskirchen R, Krautler B, Bister K.
Solution structure of the carboxyl-terminal LIM domain from quail cysteine-rich protein
CRP2.
J Biol Chem. 1997 May 2;272(18):12001-7.

Solution structure of the carboxyl-terminal LIM domain from quail cysteine-rich protein CRP2.

Members of the cysteine- and glycine-rich protein family (CRP1, CRP2, and CRP3)
contain two zinc-binding LIM domains, LIM1 (amino-terminal) and LIM2
(carboxyl-terminal), and are implicated in diverse cellular processes linked to
differentiation, growth control, and pathogenesis. Here we report the solution
structure of full-length recombinant quail CRP2 as determined by
multi-dimensional triple-resonance NMR spectroscopy. The structural analysis
revealed that the global fold of the two LIM domains in the context of the
full-length protein is identical to the recently determined solution structures
of the isolated individual LIM domains of quail CRP2. There is no preference in
relative spatial orientation of the two domains. This supports the view that
the two LIM domains are independent structural and presumably functional
modules of CRP proteins. This is also reflected by the dynamic properties of
CRP2 probed by 15N relaxation values (T1, T2, and nuclear Overhauser effect). A
model-free analysis revealed local variations in mobility along the backbone of
the two LIM domains in the native protein, similar to those observed for the
isolated domains. Interestingly, fast and slow motions observed in the 58-amino
acid linker region between the two LIM domains endow extensive motional freedom
to CRP2. The dynamic analysis indicates independent backbone mobility of the
two LIM domains and rules out correlated LIM domain motion in full-length CRP2.
The finding that the LIM domains in a protein encompassing multiple LIM motifs
are structurally and dynamically independent from each other supports the
notion that these proteins may function as adaptor molecules arranging two or
more protein constituents into a macromolecular complex.

Konrat R, Krautler B, Weiskirchen R, Bister K.
Structure of cysteine- and glycine-rich protein CRP2. Backbone dynamics reveal motional
freedom and independent spatial orientation of the lim domains.
J Biol Chem. 1998 Sep 4;273(36):23233-40.

Structure of cysteine- and glycine-rich protein CRP2. Backbone dynamics reveal motional freedom and independent spatial orientation of the lim domains.

Quail Cysteine And Glycine-Rich Protein, Nmr, Minimized Average Structure

Mutant R122a Of Quail Cysteine And Glycine-Rich Protein, Nmr, Minimized Structure