PMID- 30624066
OWN - NLM
STAT- MEDLINE
DCOM- 20200810
LR  - 20211204
IS  - 1520-6882 (Electronic)
IS  - 0003-2700 (Linking)
VI  - 91
IP  - 3
DP  - 2019 Feb 5
TI  - Multipronged ESI-MS Approach for Studying Glycan-Binding Protein Interactions 
      with Glycoproteins.
PG  - 2140-2147
LID - 10.1021/acs.analchem.8b04673 [doi]
AB  - A multipronged electrospray ionization mass spectrometry (ESI-MS) approach for 
      investigating glycan-mediated interactions between water-soluble glycan-binding 
      proteins (GBPs) and glycoproteins (GPs) is described. First, the 
      catch-and-release (CaR)-ESI-MS assay, carried with ion mobility separation prior 
      to GBP "release" (i.e., CaR(IMS)-ESI-MS), is employed to rapidly identify GBP-GP 
      binding in solution. The apparent affinity ( K(a)) of the GBP for the GP is then 
      measured using the competitive proxy ligand-ESI-MS binding assay. Finally, 
      N-glycans, enzymatically released as free oligosaccharides from the GP, are 
      screened against the GBP using ESI-MS to identify the glycans that are recognized 
      by the GBP. Measurements performed at multiple GBP concentrations allow for the 
      affinities of released N-glycans (grouped as compositional isomers) to be ranked. 
      Implementation of the approach is illustrated using the known interactions 
      between a C-terminal domain fragment of human galectin-3 (hGal-3C) and three 
      human serum GPs, alpha-1-acid glycoprotein (AGP), haptoglobin phenotype 1-1 (Hp1-1) 
      and alpha-2-macroglobulin (alpha2M). Specific binding of hGal-3C to each GP was 
      successfully detected by CaR(IMS)-ESI-MS; no binding was detected for a 
      noninteracting reference protein, which served as a negative control. The K(a) 
      measured by proxy ligand-ESI-MS for AGP, Hp1-1 and alpha2M (4 x 10(5) M(-1), 2 x 
      10(5) M(-1) and 3 x 10(5) M(-1), respectively) are consistent with the reported 
      apparent affinities of hGal-3 for serum GPs and their N-glycans. Screening the 
      N-glycan libraries of each of the GPs against hGal-3C identified ligands 
      corresponding to a total of 20 different saccharide compositions with sialylated 
      bi-, tri-, and tetra-antennary structures. The results of binding measurements 
      performed at two different hGal-3C concentrations revealed that all of the 
      N-glycan ligands exhibit affinities >/=10(4) M(-1).
FAU - Wang, Yilin
AU  - Wang Y
AD  - Alberta Glycomics Centre and Department of Chemistry , University of Alberta , 
      Edmonton , Alberta T6G 2G2 , Canada.
FAU - Park, Heajin
AU  - Park H
AD  - Alberta Glycomics Centre and Department of Chemistry , University of Alberta , 
      Edmonton , Alberta T6G 2G2 , Canada.
FAU - Lin, Hong
AU  - Lin H
AD  - Alberta Glycomics Centre and Department of Chemistry , University of Alberta , 
      Edmonton , Alberta T6G 2G2 , Canada.
FAU - Kitova, Elena N
AU  - Kitova EN
AD  - Alberta Glycomics Centre and Department of Chemistry , University of Alberta , 
      Edmonton , Alberta T6G 2G2 , Canada.
FAU - Klassen, John S
AU  - Klassen JS
AUID- ORCID: 0000-0002-3389-7112
AD  - Alberta Glycomics Centre and Department of Chemistry , University of Alberta , 
      Edmonton , Alberta T6G 2G2 , Canada.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20190125
PL  - United States
TA  - Anal Chem
JT  - Analytical chemistry
JID - 0370536
RN  - 0 (Blood Proteins)
RN  - 0 (Galectin 3)
RN  - 0 (Galectins)
RN  - 0 (Glycoproteins)
RN  - 0 (LGALS3 protein, human)
RN  - 0 (Ligands)
RN  - 0 (Polysaccharides)
SB  - IM
MH  - Blood Proteins
MH  - Galectin 3/blood/*chemistry
MH  - Galectins
MH  - Glycoproteins/blood/*chemistry
MH  - Humans
MH  - Ligands
MH  - Polysaccharides/blood/chemistry
MH  - Protein Binding
MH  - Spectrometry, Mass, Electrospray Ionization
EDAT- 2019/01/10 06:00
MHDA- 2020/08/11 06:00
CRDT- 2019/01/10 06:00
PHST- 2019/01/10 06:00 [pubmed]
PHST- 2020/08/11 06:00 [medline]
PHST- 2019/01/10 06:00 [entrez]
AID - 10.1021/acs.analchem.8b04673 [doi]
PST - ppublish
SO  - Anal Chem. 2019 Feb 5;91(3):2140-2147. doi: 10.1021/acs.analchem.8b04673. Epub 
      2019 Jan 25.