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Glycoproteomic identification of galectin-3 and -8 ligands in bronchoalveolar lavage of mild asthmatics and healthy subjects.

Cederfur, Cecilia LU ; Malmström, Johan ; Nihlberg, Kristian LU ; Block, Mattias ; Breimer, Michael E ; Bjermer, Leif LU ; Westergren-Thorsson, Gunilla LU and Leffler, Hakon LU (2012) In Biochimica et Biophysica Acta 1820(9). p.1429-1436
Abstract
BACKGROUND: Galectins, a family of small carbohydrate binding proteins, have been implicated in regulation of inflammatory reactions, including asthma and fibrosis in the lungs. Galectins are found in cells of the airways and in airway secretions, but their glycoprotein ligands there have only been studied to a very limited extent. METHODS: Bronchoalveolar lavage (BAL) fluid from mild asthmatics and healthy volunteers were fractionated by affinity chromatography on the immobilized galectins. Total (10-30μg) and galectin bound (~1-10μg) protein fractions were identified, quantified and compared using shot-gun proteomics and spectral counts. RESULTS: About 175 proteins were identified in unfractionated BAL-fluid, and about 100 bound... (More)
BACKGROUND: Galectins, a family of small carbohydrate binding proteins, have been implicated in regulation of inflammatory reactions, including asthma and fibrosis in the lungs. Galectins are found in cells of the airways and in airway secretions, but their glycoprotein ligands there have only been studied to a very limited extent. METHODS: Bronchoalveolar lavage (BAL) fluid from mild asthmatics and healthy volunteers were fractionated by affinity chromatography on the immobilized galectins. Total (10-30μg) and galectin bound (~1-10μg) protein fractions were identified, quantified and compared using shot-gun proteomics and spectral counts. RESULTS: About 175 proteins were identified in unfractionated BAL-fluid, and about 100 bound galectin-3 and 60 bound galectin-8. These included plasma glycoproteins, and typical airway proteins such as SP-A2, PIGR and SP-B. The concentration of galectin-binding proteins was 100-300 times higher than the concentration of galectins in BAL. CONCLUSION: The low relative concentration of galectins in BAL makes it likely that functional interactions with glycoproteins occur at sites rich in galectin, such as cells of the airways, rather than the extracellular fluid itself. The profile of galectin bound proteins differed between samples from asthma patients and healthy subjects and correlated with the presence of fibroblasts or eosinophils. This included appearance of a specific galectin-8-binding glycoform of haptoglobin, previously shown to be increased in serum in other inflammatory conditions. GENERAL SIGNIFICANCE: It is technically feasible to identify galectin-binding glycoproteins in low concentration patient samples such as BAL-fluid, to generate biomedically interesting results. This article is part of a Special Issue entitled Glycoproteomics. (Less)
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organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Biochimica et Biophysica Acta
volume
1820
issue
9
pages
1429 - 1436
publisher
Elsevier
external identifiers
  • wos:000306444000016
  • pmid:22240167
  • scopus:84862890114
  • pmid:22240167
ISSN
0006-3002
DOI
10.1016/j.bbagen.2011.12.016
language
English
LU publication?
yes
additional info
The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Stem Cell Aging (013212073), Respiratory Medicine and Allergology (013230111), Lung Biology (013212002), Division of Microbiology, Immunology and Glycobiology - MIG (013025200), Neuronano Research Center (NRC) (013210020)
id
4a203980-34b8-4e2d-b976-d460a3760817 (old id 2336461)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/22240167?dopt=Abstract
date added to LUP
2016-04-01 14:38:39
date last changed
2022-01-28 01:46:59
@article{4a203980-34b8-4e2d-b976-d460a3760817,
  abstract     = {{BACKGROUND: Galectins, a family of small carbohydrate binding proteins, have been implicated in regulation of inflammatory reactions, including asthma and fibrosis in the lungs. Galectins are found in cells of the airways and in airway secretions, but their glycoprotein ligands there have only been studied to a very limited extent. METHODS: Bronchoalveolar lavage (BAL) fluid from mild asthmatics and healthy volunteers were fractionated by affinity chromatography on the immobilized galectins. Total (10-30μg) and galectin bound (~1-10μg) protein fractions were identified, quantified and compared using shot-gun proteomics and spectral counts. RESULTS: About 175 proteins were identified in unfractionated BAL-fluid, and about 100 bound galectin-3 and 60 bound galectin-8. These included plasma glycoproteins, and typical airway proteins such as SP-A2, PIGR and SP-B. The concentration of galectin-binding proteins was 100-300 times higher than the concentration of galectins in BAL. CONCLUSION: The low relative concentration of galectins in BAL makes it likely that functional interactions with glycoproteins occur at sites rich in galectin, such as cells of the airways, rather than the extracellular fluid itself. The profile of galectin bound proteins differed between samples from asthma patients and healthy subjects and correlated with the presence of fibroblasts or eosinophils. This included appearance of a specific galectin-8-binding glycoform of haptoglobin, previously shown to be increased in serum in other inflammatory conditions. GENERAL SIGNIFICANCE: It is technically feasible to identify galectin-binding glycoproteins in low concentration patient samples such as BAL-fluid, to generate biomedically interesting results. This article is part of a Special Issue entitled Glycoproteomics.}},
  author       = {{Cederfur, Cecilia and Malmström, Johan and Nihlberg, Kristian and Block, Mattias and Breimer, Michael E and Bjermer, Leif and Westergren-Thorsson, Gunilla and Leffler, Hakon}},
  issn         = {{0006-3002}},
  language     = {{eng}},
  number       = {{9}},
  pages        = {{1429--1436}},
  publisher    = {{Elsevier}},
  series       = {{Biochimica et Biophysica Acta}},
  title        = {{Glycoproteomic identification of galectin-3 and -8 ligands in bronchoalveolar lavage of mild asthmatics and healthy subjects.}},
  url          = {{http://dx.doi.org/10.1016/j.bbagen.2011.12.016}},
  doi          = {{10.1016/j.bbagen.2011.12.016}},
  volume       = {{1820}},
  year         = {{2012}},
}