Galectin-1-binding glycoforms of haptoglobin with altered intracellular trafficking, and increase in metastatic breast cancer patients.

Abstract

Sera from 25 metastatic breast cancer patients and 25 healthy controls were subjected to affinity chromatography using immobilized galectin-1. Serum from the healthy subjects contained on average 1.2 mg per ml (range 0.7-2.2) galectin-1 binding glycoproteins, whereas serum from the breast cancer patients contained on average 2.2 mg/ml (range 0.8-3.9), with a higher average for large primary tumours. The major bound glycoproteins were α-2-macroglobulin, IgM and haptoglobin. Both the IgM and haptoglobin concentrations were similar in cancer compared to control sera, but the percentage bound to galectin-1 was lower for IgM and higher for haptoglobin: about 50% (range 20-80) in cancer sera and about 30% (range 25-50) in healthy sera. Galectin-1 binding and non-binding fractions were separated by affinity chromatography from pooled haptoglobin from healthy sera. The N-glycans of each fraction were analyzed by mass spectrometry, and the structural differences and galectin-1 mutants were used to identify possible galectin-1 binding sites. Galectin-1 binding and non-binding fractions were also analyzed regarding their haptoglobin function. Both were similar in forming complex with haemoglobin and mediate its uptake into alternatively activated macrophages. However, after uptake there was a dramatic difference in intracellular targeting, with the galectin-1 non-binding fraction going to a LAMP-2 positive compartment (lysosomes), while the galectin-1 binding fraction went to larger galectin-1 positive granules. In conclusion, galectin-1 detects a new type of functional biomarker for cancer: a specific type of glycoform of haptoglobin, and possibly other serum glycoproteins, with a different function after uptake into tissue cells.

• Authors: Michael C Carlsson, Cecilia Cederfur, Viveka Schaar, Crina I A Balog, Adriana Lepur, Franck Touret, Emma Salomonsson, André M Deelder, Mårten Fernö, Håkan Olsson, Manfred Wuhrer, Hakon Leffler
• Journal: PloS one (PLoS One) Vol. 6 Issue 10 Pg. e26560 ( 2011) ISSN: 1932-6203 [Electronic] United States
• PMID: 22028908 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Galectin 1
• Haptoglobins
• Immobilized Proteins
• Immunoglobulin M
• Polysaccharides
• N-Acetylneuraminic Acid

Topics

• Aged
• Binding Sites
• Breast Neoplasms (metabolism, pathology)
• Case-Control Studies
• Cell Line, Tumor
• Endocytosis
• Female
• Galectin 1 (chemistry, immunology, metabolism)
• Haptoglobins (chemistry, metabolism)
• Humans
• Immobilized Proteins (chemistry, metabolism)
• Immunoglobulin M (immunology)
• Intracellular Space (metabolism)
• Macrophages (cytology, metabolism)
• Middle Aged
• Models, Molecular
• N-Acetylneuraminic Acid
• Neoplasm Metastasis
• Polysaccharides (metabolism)
• Protein Binding
• Protein Multimerization
• Protein Structure, Quaternary
• Protein Transport
• Substrate Specificity