PMID- 27801570
OWN - NLM
STAT- MEDLINE
DCOM- 20180822
LR  - 20220403
IS  - 1520-6882 (Electronic)
IS  - 0003-2700 (Linking)
VI  - 88
IP  - 23
DP  - 2016 Dec 6
TI  - Enrichment of Two Isomeric Heparin Oligosaccharides Exhibiting Different 
      Affinities toward Monocyte Chemoattractant Protein-1.
PG  - 11551-11558
AB  - Chemokine-GAG interactions are crucial to facilitate chemokine immobilization, 
      resulting in the formation of chemokine gradients that guide cell migration. Here 
      we demonstrate chromatographic isolation and purification of two heparin 
      hexasaccharide isomers that interact with the oligomeric chemokine Monocyte 
      Chemoattractant Protein-1 (MCP-1)/CCL2 with different binding affinities. The 
      sequences of these two hexasaccharides were deduced from unique MS/MS product 
      ions and HPLC compositional analysis. Ion mobility mass spectrometry (IM-MS) 
      showed that the two isolated oligosaccharides have different conformations and 
      both displayed preferential binding for one of the two distinct conformations 
      known for MCP-1 dimers. A significant shift in arrival time distribution of close 
      to 70 A(2) was observed, indicating a more compact protein:hexasaccharide 
      conformation. Clear differences in the MS spectra between bound and unbound 
      protein allowed calculation of K(d) values from the resulting data. The 
      structural difference between the two hexasaccharides was defined as the 
      differential location of a single sulfate at either C-6 of glucosamine or C-2 of 
      uronic acid in the reducing disaccharide, resulting in a 200-fold difference in 
      binding affinity for MCP-1. These data indicate sequence specificity for high 
      affinity binding, supporting the view that sulfate position, and not simply the 
      number of sulfates, is important for heparan sulfate protein binding.
FAU - Miller, Rebecca L
AU  - Miller RL
AD  - Departments of Molecular and Cellular Biology and Chemistry, University of 
      California , 1 Shields Drive, Davis, California 95616, United States.
FAU - Dykstra, Andrew B
AU  - Dykstra AB
AD  - Departments of Molecular and Cellular Biology and Chemistry, University of 
      California , 1 Shields Drive, Davis, California 95616, United States.
FAU - Wei, Wei
AU  - Wei W
AD  - Departments of Molecular and Cellular Biology and Chemistry, University of 
      California , 1 Shields Drive, Davis, California 95616, United States.
FAU - Holsclaw, Cynthia
AU  - Holsclaw C
AD  - Departments of Molecular and Cellular Biology and Chemistry, University of 
      California , 1 Shields Drive, Davis, California 95616, United States.
FAU - Turnbull, Jeremy E
AU  - Turnbull JE
AD  - Centre for Glycobiology, Department of Biochemistry, Institute of Integrative 
      Biology, University of Liverpool , Crown Street, Liverpool, L69 7ZB, England.
FAU - Leary, Julie A
AU  - Leary JA
AD  - Departments of Molecular and Cellular Biology and Chemistry, University of 
      California , 1 Shields Drive, Davis, California 95616, United States.
LA  - eng
GR  - BB/I004343/1/BB_/Biotechnology and Biological Sciences Research Council/United 
      Kingdom
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20161115
PL  - United States
TA  - Anal Chem
JT  - Analytical chemistry
JID - 0370536
RN  - 0 (CCL2 protein, human)
RN  - 0 (Chemokine CCL2)
RN  - 0 (Oligosaccharides)
RN  - 9005-49-6 (Heparin)
SB  - IM
MH  - Chemokine CCL2/*analysis
MH  - Chromatography, High Pressure Liquid
MH  - Heparin/*chemistry
MH  - Humans
MH  - Isomerism
MH  - Oligosaccharides/*chemistry
MH  - Tandem Mass Spectrometry
EDAT- 2016/11/02 06:00
MHDA- 2018/08/23 06:00
CRDT- 2016/11/02 06:00
PHST- 2016/11/02 06:00 [pubmed]
PHST- 2018/08/23 06:00 [medline]
PHST- 2016/11/02 06:00 [entrez]
AID - 10.1021/acs.analchem.6b02803 [doi]
PST - ppublish
SO  - Anal Chem. 2016 Dec 6;88(23):11551-11558. doi: 10.1021/acs.analchem.6b02803. Epub 
      2016 Nov 15.