NMR structure of human I-type lectin domain-glycan complex

human I-type lectin domain-glycan complex

[(2R,3R,4S,5R,6R)-3,4,5,6-tetrahydroxyoxan-2-yl]methyl hydrogen sulfate

(2R,4S,5R,6R)-5-acetamido-2,4-dihydroxy-6-[(1R,2R)-1,2,3-trihydroxypropyl]oxane-2-carboxylic acid

5-(acetylamino)-3,5-dideoxy-D-glycero-alpha-D-galacto-non-2-ulopyranosonic acid

Proceedings of the National Academy of Sciences of the United States of America
Structural basis for sulfation-dependent self-glycan recognition by the human immune-inhibitory receptor Siglec-8.

numbering corresponds to mature protein sequence
Construct includes Siglec-8 residues M1-H139 (carrying a C26S point mutation to enable soluble expression; numbering corresponds to mature protein), followed by six non-native C-terminal residues G140-R145 (remainder of the thrombin cleavage site after affinity-tag removal)

Neu5Ac_alpha2-3[6O-SO3-]Gal_beta1-4(Fuc_alpha1-3)GlcNAc_beta_spacer
//Ligand = 6-"prime"-sulfo-sialyl Lewis X tetrasaccharide with artificial chemical linker/spacer (SP) (attached to NAG C1-atom)

Insert encoding for Siglec-8 lectin domain including residues M1-H139 (carrying C26S point mutation).
Construct design described in:
Proepster, J.M., et al, Protein Expr. Purif. 2015, 109C: 14-22

6"prime"-sulfo sialyl Lewis X-O-(3-amino)propyl glycoside:
Neu5Ac alpha2-3[6O-SO3-]Gal beta1-4(Fuc alpha1-3)GlcNAc-beta-O(CH2)3NH3+

Full-lenght cDNA clone: IRAUp969G05104D(imaGenes GmbH, Germany);

NMR structure of human I-type lectin domain
simulated annealing and automated NOE assignment
Sialic acid-binding immunoglobulin-like lectin 8
protein backbone torsion angle prediction from NMR chemical shifts
Case, Darden, Cheatham, III, Simmerling, Wang, Duke, Luo, ... and Kollman