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@ARTICLE{Boix:144054,
author = {Boix, E. and Pulido, D. and Moussaoui, M. and Nogues, M. V.
and Russi, S. and DESY},
title = {{T}he sulfate-binding site structure of the human
eosinophil cationic protein as revealed by a new crystal
form},
journal = {Journal of structural biology},
volume = {179},
issn = {1047-8477},
address = {San Diego, Calif.},
publisher = {Elsevier},
reportid = {PHPPUBDB-25783},
pages = {1-9},
year = {2012},
abstract = {The human eosinophil cationic protein (ECP), also known as
RNase 3, is an eosinophil secretion protein that is involved
in innate immunity and displays antipathogen and
proinflammatory activities. ECP has a high binding affinity
for heterosaccharides, such as bacterial lipopolysaccharides
and heparan sulfate found in the glycocalix of eukaryotic
cells. We have crystallized ECP in complex with sulfate
anions in a new monoclinic crystal form. In this form, the
active site groove is exposed, providing an alternative
model for ligand binding studies. By exploring the
protein-sulfate complex, we have defined the sulfate binding
site architecture. Three main sites (S1-S3) are located in
the protein active site; S1 and S2 overlap with the
phosphate binding sites involved in RNase nucleotide
recognition. A new site (S3) that is unique to ECP is one of
the key anchoring points for sulfated ligands. Arg 1 and Arg
7 in S3, together with Arg 34 and Arg 36 in S1, form the
main basic clusters that assist in the recognition of ligand
anionic groups. The location of additional sulfate bound
molecules, some of which contribute to the crystal packing,
may mimic the binding to extended anionic polymers. In
conclusion, the structural data define a binding pattern for
the recognition of sulfated molecules that can modulate the
role of ECP in innate immunity. The results reveal the
structural basis for the high affinity of ECP for
glycosaminoglycans and can assist in structure-based drug
design of inhibitors of the protein cytotoxicity to host
tissues during inflammation.},
keywords = {Amino Acid Sequence / Catalytic Domain / Crystallization /
Eosinophil Cationic Protein: chemistry / Eosinophil Cationic
Protein: metabolism / Glycosaminoglycans: chemistry /
Glycosaminoglycans: metabolism / Humans / Ligands /
Molecular Dynamics Simulation / Molecular Sequence Data /
Protein Binding / Protein Conformation / Recombinant
Proteins: chemistry / Recombinant Proteins: metabolism /
Sulfates: chemistry / Glycosaminoglycans (NLM Chemicals) /
Ligands (NLM Chemicals) / Recombinant Proteins (NLM
Chemicals) / Sulfates (NLM Chemicals) / Eosinophil Cationic
Protein (NLM Chemicals)},
cin = {HASYLAB},
ddc = {540},
cid = {$I:(DE-H253)HASYLAB_-2012_-20130307$},
pnm = {DORIS Beamline K1.1 (POF2-54G13)},
pid = {G:(DE-H253)POF2-K1.1-20130405},
experiment = {EXP:(DE-H253)D-K1.1-20150101},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:22579681},
UT = {WOS:000305775400001},
doi = {10.1016/j.jsb.2012.04.023},
url = {https://bib-pubdb1.desy.de/record/144054},
}