PMID- 25536525
OWN - NLM
STAT- MEDLINE
DCOM- 20151208
LR  - 20150205
IS  - 1477-0539 (Electronic)
IS  - 1477-0520 (Linking)
VI  - 13
IP  - 7
DP  - 2015 Feb 21
TI  - Phosphate modulates receptor sulfotyrosine recognition by the chemokine monocyte 
      chemoattractant protein-1 (MCP-1/CCL2).
PG  - 2162-9
LID - 10.1039/c4ob02262a [doi]
AB  - Tyrosine sulfation is a widespread post-translational modification that mediates 
      the interactions of secreted and membrane-associated proteins in such varied 
      biological processes as peptide hormone action, adhesion, blood coagulation, 
      complement activation and regulation of leukocyte trafficking. Due to the 
      heterogeneous nature of tyrosine sulfation, detailed biochemical and biophysical 
      studies of tyrosine sulfation rely on homogenous, synthetic sulfopeptides. Here 
      we describe the synthesis of a fluorescent sulfopeptide (FL-R2D) derived from the 
      chemokine receptor CCR2 and the application of FL-R2D in direct and competitive 
      fluorescence anisotropy assays that enable the efficient measurement of binding 
      affinities between sulfopeptides and their binding proteins. Using these assays, 
      we have found that the binding of the chemokine monocyte chemoattractant 
      protein-1 (MCP-1) to sulfated peptides derived from the chemokine receptor CCR2 
      is highly dependent on the assay buffer. In particular, phosphate buffer at close 
      to physiological concentrations competes with the receptor sulfopeptide by 
      binding to the sulfopeptide binding pocket on the chemokine surface. Thus, 
      physiological phosphate may modulate the receptor binding selectivity of 
      chemokines.
FAU - Ludeman, Justin P
AU  - Ludeman JP
AD  - Department of Biochemistry and Molecular Biology, Monash University, Clayton, VIC 
      3800, Australia. martin.stone@monash.edu.
FAU - Nazari-Robati, Mahdieh
AU  - Nazari-Robati M
FAU - Wilkinson, Brendan L
AU  - Wilkinson BL
FAU - Huang, Cheng
AU  - Huang C
FAU - Payne, Richard J
AU  - Payne RJ
FAU - Stone, Martin J
AU  - Stone MJ
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Org Biomol Chem
JT  - Organic & biomolecular chemistry
JID - 101154995
RN  - 0 (CCL2 protein, human)
RN  - 0 (Chemokine CCL2)
RN  - 0 (Phosphates)
RN  - 29166358BF (tyrosine O-sulfate)
RN  - 42HK56048U (Tyrosine)
SB  - IM
MH  - Binding Sites
MH  - Chemokine CCL2/chemistry/*metabolism
MH  - Molecular Conformation
MH  - Phosphates/chemistry/*metabolism
MH  - Tyrosine/*analogs & derivatives/chemistry/metabolism
EDAT- 2014/12/24 06:00
MHDA- 2015/12/15 06:00
CRDT- 2014/12/24 06:00
PHST- 2014/12/24 06:00 [entrez]
PHST- 2014/12/24 06:00 [pubmed]
PHST- 2015/12/15 06:00 [medline]
AID - 10.1039/c4ob02262a [doi]
PST - ppublish
SO  - Org Biomol Chem. 2015 Feb 21;13(7):2162-9. doi: 10.1039/c4ob02262a.