Identification and Characterization of a Glycosaminoglycan Recognition Element of the C Chemokine Lymphotactin

Francis C. Peterson, E. Sonay Elgin, Timothy J Nelson, Fuming Zhang, Theresa J. Hoeger, Robert J. Linhardt, Brian F. Volkman

Research output: Contribution to journalArticle

60 Citations (Scopus)

Abstract

Chemokine-mediated recruitment of leukocytes in vivo depends on interactions with cell surface glycosaminoglycans. Lymphotactin, the unique member of the "C" chemokine subclass, is a highly basic protein that binds heparin, a glycosaminoglycan, with high affinity (∼10 nM). We detected lymphotactin-heparin binding by NMR and mapped this interaction to a narrow surface that wraps around the protein. Substitutions in and around this binding site and surface plasmon resonance analysis of heparin binding affinity identified two arginine residues of lymphotactin as critical for glycosaminoglycan binding. Both arginine mutant proteins and the combined double mutant had dramatically diminished in vivo activity in a leukocyte recruitment assay, suggesting that the lymphotactin-glycosaminoglycan interactions detected in vitro are important for the function of this chemokine. Our results demonstrate that like other chemokines, lymphotactin utilizes highly specific glycosaminoglycan-binding sites that represent potential targets for drug development.

Original languageEnglish (US)
Pages (from-to)12598-12604
Number of pages7
JournalJournal of Biological Chemistry
Volume279
Issue number13
DOIs
StatePublished - Mar 26 2004
Externally publishedYes

Fingerprint

C Chemokines
Glycosaminoglycans
Chemokines
Heparin
Arginine
Leukocytes
Binding Sites
Surface Plasmon Resonance
Surface plasmon resonance
Mutant Proteins
Cell Communication
Assays
Proteins
Substitution reactions
Nuclear magnetic resonance
lymphotactin
Pharmaceutical Preparations

ASJC Scopus subject areas

  • Biochemistry

Cite this

Identification and Characterization of a Glycosaminoglycan Recognition Element of the C Chemokine Lymphotactin. / Peterson, Francis C.; Elgin, E. Sonay; Nelson, Timothy J; Zhang, Fuming; Hoeger, Theresa J.; Linhardt, Robert J.; Volkman, Brian F.

In: Journal of Biological Chemistry, Vol. 279, No. 13, 26.03.2004, p. 12598-12604.

Research output: Contribution to journalArticle

Peterson, Francis C. ; Elgin, E. Sonay ; Nelson, Timothy J ; Zhang, Fuming ; Hoeger, Theresa J. ; Linhardt, Robert J. ; Volkman, Brian F. / Identification and Characterization of a Glycosaminoglycan Recognition Element of the C Chemokine Lymphotactin. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 13. pp. 12598-12604.
@article{779162f93ec84edbafb3017952305f80,
title = "Identification and Characterization of a Glycosaminoglycan Recognition Element of the C Chemokine Lymphotactin",
abstract = "Chemokine-mediated recruitment of leukocytes in vivo depends on interactions with cell surface glycosaminoglycans. Lymphotactin, the unique member of the {"}C{"} chemokine subclass, is a highly basic protein that binds heparin, a glycosaminoglycan, with high affinity (∼10 nM). We detected lymphotactin-heparin binding by NMR and mapped this interaction to a narrow surface that wraps around the protein. Substitutions in and around this binding site and surface plasmon resonance analysis of heparin binding affinity identified two arginine residues of lymphotactin as critical for glycosaminoglycan binding. Both arginine mutant proteins and the combined double mutant had dramatically diminished in vivo activity in a leukocyte recruitment assay, suggesting that the lymphotactin-glycosaminoglycan interactions detected in vitro are important for the function of this chemokine. Our results demonstrate that like other chemokines, lymphotactin utilizes highly specific glycosaminoglycan-binding sites that represent potential targets for drug development.",
author = "Peterson, {Francis C.} and Elgin, {E. Sonay} and Nelson, {Timothy J} and Fuming Zhang and Hoeger, {Theresa J.} and Linhardt, {Robert J.} and Volkman, {Brian F.}",
year = "2004",
month = "3",
day = "26",
doi = "10.1074/jbc.M311633200",
language = "English (US)",
volume = "279",
pages = "12598--12604",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "13",

}

TY - JOUR

T1 - Identification and Characterization of a Glycosaminoglycan Recognition Element of the C Chemokine Lymphotactin

AU - Peterson, Francis C.

AU - Elgin, E. Sonay

AU - Nelson, Timothy J

AU - Zhang, Fuming

AU - Hoeger, Theresa J.

AU - Linhardt, Robert J.

AU - Volkman, Brian F.

PY - 2004/3/26

Y1 - 2004/3/26

N2 - Chemokine-mediated recruitment of leukocytes in vivo depends on interactions with cell surface glycosaminoglycans. Lymphotactin, the unique member of the "C" chemokine subclass, is a highly basic protein that binds heparin, a glycosaminoglycan, with high affinity (∼10 nM). We detected lymphotactin-heparin binding by NMR and mapped this interaction to a narrow surface that wraps around the protein. Substitutions in and around this binding site and surface plasmon resonance analysis of heparin binding affinity identified two arginine residues of lymphotactin as critical for glycosaminoglycan binding. Both arginine mutant proteins and the combined double mutant had dramatically diminished in vivo activity in a leukocyte recruitment assay, suggesting that the lymphotactin-glycosaminoglycan interactions detected in vitro are important for the function of this chemokine. Our results demonstrate that like other chemokines, lymphotactin utilizes highly specific glycosaminoglycan-binding sites that represent potential targets for drug development.

AB - Chemokine-mediated recruitment of leukocytes in vivo depends on interactions with cell surface glycosaminoglycans. Lymphotactin, the unique member of the "C" chemokine subclass, is a highly basic protein that binds heparin, a glycosaminoglycan, with high affinity (∼10 nM). We detected lymphotactin-heparin binding by NMR and mapped this interaction to a narrow surface that wraps around the protein. Substitutions in and around this binding site and surface plasmon resonance analysis of heparin binding affinity identified two arginine residues of lymphotactin as critical for glycosaminoglycan binding. Both arginine mutant proteins and the combined double mutant had dramatically diminished in vivo activity in a leukocyte recruitment assay, suggesting that the lymphotactin-glycosaminoglycan interactions detected in vitro are important for the function of this chemokine. Our results demonstrate that like other chemokines, lymphotactin utilizes highly specific glycosaminoglycan-binding sites that represent potential targets for drug development.

UR - http://www.scopus.com/inward/record.url?scp=1842477451&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=1842477451&partnerID=8YFLogxK

U2 - 10.1074/jbc.M311633200

DO - 10.1074/jbc.M311633200

M3 - Article

VL - 279

SP - 12598

EP - 12604

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 13

ER -