Lysines and Arginines play non-redundant roles in mediating chemokine-glycosaminoglycan interactions

Prem Raj B. Joseph, Kirti V. Sawant, Junji Iwahara, Roberto Garofalo, Umesh R. Desai, Krishna Rajarathnam

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Glycosaminoglycans (GAGs) bind a large array of proteins and mediate fundamental and diverse roles in human physiology. Ion pair interactions between protein lysines/arginines and GAG sulfates/carboxylates mediate binding. Neutrophil-activating chemokines (NAC) are GAG-binding proteins, and their sequences reveal high selectivity for lysines over arginines indicating they are functionally not equivalent. NAC binding to GAGs impacts gradient formation, receptor functions, and endothelial activation, which together regulate different components of neutrophil migration. We characterized the consequence of mutating lysine to arginine in NAC CXCL8, a well-characterized GAG-binding protein. We chose three lysines — two highly conserved lysines (K20 and K64) and a CXCL8-specific lysine (K67). Interestingly, the double K64R/K20R and K64R/K67R mutants are highly impaired in recruiting neutrophils in a mouse model. Further, both the mutants bind GAG heparin with higher affinity but show similar receptor activity. NMR and MD studies indicate that the structures are essentially identical to the WT, but the mutations alter the network of intramolecular ion pair interactions. These observations collectively indicate that the reduced in vivo recruitment is due to altered GAG interactions, higher GAG binding affinity can be detrimental, and specificity of lysines fine-tunes in vivo GAG interactions and function.

Original languageEnglish (US)
Article number12289
JournalScientific Reports
Volume8
Issue number1
DOIs
StatePublished - Dec 1 2018

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Glycosaminoglycans
Chemokines
Lysine
Arginine
Carrier Proteins
Ions
Physiology
Interleukin-8
Sulfates
Heparin
Proteins
Chemical activation
Nuclear magnetic resonance

ASJC Scopus subject areas

  • General

Cite this

Lysines and Arginines play non-redundant roles in mediating chemokine-glycosaminoglycan interactions. / Joseph, Prem Raj B.; Sawant, Kirti V.; Iwahara, Junji; Garofalo, Roberto; Desai, Umesh R.; Rajarathnam, Krishna.

In: Scientific Reports, Vol. 8, No. 1, 12289, 01.12.2018.

Research output: Contribution to journalArticle

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