Lymphotactin, the sole identified member of the C class of chemokines, spec
ifically attracts T lymphocytes and natural killer cells. This 93-residue p
rotein lacks 2 of the 4 conserved cysteine residues characteristic of the o
ther 3 classes of chemokines and possesses an extended carboxyl terminus, w
hich is required for chemotactic activity. We have determined the three-dim
ensional solution structure of,recombinant human lymphotactin by NMR spectr
oscopy. Under the conditions used for the structure determination, lymphota
ctin was predominantly monomeric; however, pulsed field gradient NMR selfdi
ffusion measurements and analytical ultracentrifugation revealed evidence o
f dimer formation. Sequence-specific chemical shift assignments were determ
ined through analysis of two- and three-dimensional NMR spectra of N-15- an
d C-13/N-15-enriched protein samples. Input for the torsion angle dynamics
calculations used in determining the structure included 1258 unique NOE-der
ived distance constraints and 60 dihedral angle constraints obtained from c
hemical-shift-based searching of a protein conformational database. The ens
emble of 20 structures chosen to represent the structure had backbone and h
eavy atom rms deviations of 0.46 +/- 0.11 and 1.02 +/- 0.14 Angstrom, respe
ctively. The results revealed that human lymphotactin adopts the conserved
chemokine fold, which is characterized by a three-stranded antiparallel bet
a -sheet and a C-terminal a-helix. Two regions are dynamically disordered a
s evidenced by H-1 and C-13 Chemical shifts and {N-15}- NOES: residues 1-9
of the amino terminus and residues 69-93 of the C-terminal extension. A fun
ctional role for the C-terminal extension, which is unique to lymphotactin,
remains to be elucidated.