1H, 15N and 13C Assignments of FLIN2, an Intramolecular LMO2:ldb1 Complex

Intramolecular complex of LMO2(LIM1) and ldb1(LID)

Ldb1 is a nuclear protein that binds to a number of nuclear LIM-domain containing proteins. It binds to LMO2 and this binary complex is thought to take part in higher order transcriptional complexes.

LMO2 (otherwise known as rhombotin-2) is an essential haematopoietic transcription factor. Its overexpression in T cells causes the onset of Acute T-cell Lymphoblastic Leukemia (T-ALL).

Some minor abundance peaks were observed that may correspond to a minor
conformational species.
The number and intensity of some of those peaks increases
with the age of the sample.
The C-terminal residue has a both a major peak (assigned) and a minor
peak (unassigned in these data sets) that appear in only some spectra.

Some minor abundance peaks were observed that may correspond to a minor
conformational species.

Information obtained from PDB's Chemical Component Dictionary
at http://wwpdb-remediation.rutgers.edu/downloads.html
Downloaded on Thu Jul 21 09:32:42 2011

The nuclear LIM-only (LMO) transcription factors LMO2 and LMO4 play important
roles in both normal and leukemic T-cell development. LIM domains are
cysteine/histidine-rich domains that contain two structural zinc ions and that
function as protein-protein adaptors; members of the LMO family each contain
two closely spaced LIM domains. These LMO proteins all bind with high affinity
to the nuclear protein LIM domain binding protein 1 (ldb1). The LMO-ldb1
interaction is mediated through the N-terminal LIM domain (LIM1) of LMO
proteins and a 38-residue region towards the C-terminus of ldb1 [ldb1(LID)].
Unfortunately, recombinant forms of LMO2 and LMO4 have limited solubility and
stability, effectively preventing structural analysis. Therefore, we have
designed and constructed a fusion protein in which ldb1(LID) and LIM1 of LMO2
can form an intramolecular complex. The engineered protein, FLIN2 (fusion of
the LIM interacting domain of ldb1 and the N-terminal LIM domain of LMO2) has
been expressed and purified in milligram quantities. FLIN2 is monomeric,
contains significant levels of secondary structure and yields a sharp and
well-dispersed one-dimensional (1)H NMR spectrum. The analogous LMO4 protein,
FLIN4, has almost identical properties. These data suggest that we will be able
to obtain high-resolution structural information about the LMO-ldb1
interactions.

J.E. Deane, E. Sum, J.P. Mackay, G.J. Lindeman, J.E. Visvader & J.M. Matthews
Protein Engineering, in press (2001).

Design, production and characterization of FLIN2 and FLIN4: the engineering of intramolecular ldb1:LMO complexes.

Residue 1 stems from the polylinker region of the plasmid.
Residues 2-62 of FLIN2 correspond to residues 26-87 of LMO2.
Residues 63-74 form a linker region.
Residues 75-114 of FLIN2 correspond to residues 300-339 of ldb1.

Fusion of the LIM interaction domain of ldb1 and the N-terminal LIM domain of LMO2

Structure Of Flin2, A Complex Containing The N-Terminal Lim Domain Of Lmo2 And Ldb1-Lid

PREDICTED: similar to LIM domain only 2 (rhombotin-like 1), [Monodelphis domestica]

distance geometry, simulated annealing, molecular dynamics, automated noe assignment of ambiguous NOES