Different affinity of galectins for human serum glycoproteins: galectin-3 binds many protease inhibitors and acute phase proteins.
Here we report the first survey of galectins binding to glycoproteins of human serum. Serum was subjected to affinity chromatography using immobilized galectins, and the bound glycoproteins were analyzed by electrophoresis, Western blotting, and mass spectrometry. Galectins-3, -8, and -9 bound a much broader range of ligands in serum than previously known, galectin-1 bound less, and galectins-2, -4, and -7 bound only traces or no serum ligands. Galectin-3 bound most major glycoproteins, including alpha-2-macroglobulin and acute phase proteins such as haptoglobin. It bound only a selected minor fraction of transferrin, and bound none or little of IgG. Galectins-8 and -9 bound a similar range of glycoproteins as galectin-3, but in lower amounts, and galectin-8 had a relative preference for IgA. Galectin-1 bound mainly a fraction of alpha-2-macroglobulin and only traces of other glycoproteins. The binding of galectin-3 to serum glycoproteins requires affinity for LacNAc, since a mutant (R186S), which has lost this affinity, did not bind any serum glycoproteins. The average affinity of galectin-3 for serum glycoproteins was estimated to correspond to K(d) approximately 1-5 muM by modeling of the affinity chromatography and a fluorescence anisotropy assay. Since galectins are expressed on endothelial cells and other cells exposed to serum components, this report gives new insight into function of galectins and the role of their different fine specificity giving differential binding to the serum glycoproteins.
Cederfur
Cecilia
C
Department of Laboratory Medicine, Section MIG (Microbiology, Immunology, Glycobiology), Lund University, 223-62 Lund, Sweden. cecilia.cederfur@med.lu.se
Salomonsson
Emma
E
Nilsson
Jonas
J
Halim
Adnan
A
Oberg
Christopher T
CT
Larson
Göran
G
Nilsson
Ulf J
UJ
Leffler
Hakon
H
D000209:Acute-Phase Proteins / Q000378:metabolism*; D001665:Binding Sites; D002846:Chromatography, Affinity; D005454:Fluorescence Polarization; D037502:Galectin 3 / Q000378:metabolism*; D037161:Galectins / Q000378:metabolism*; D006023:Glycoproteins / Q000097:blood* / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008024:Ligands; D011480:Protease Inhibitors / Q000378:metabolism*
D000209:Acute-Phase Proteins; D037502:Galectin 3; D037161:Galectins; D006023:Glycoproteins; C091991:LGALS8 protein, human; C105032:LGALS9 protein, human; D008024:Ligands; D011480:Protease Inhibitors