Abstract
Galectin-3 binding protein (LGALS3BP or 90 K) is a secreted glycoprotein found in human body fluids. Deregulated levels were observed in cancer and infection and its study in neurological diseases is more recent. Here, we have investigated 90 K from human cerebrospinal fluid (CSF) of patients with amyotrophic lateral sclerosis (ALS, n = 35) and other neurological diseases (n = 23). CSF was fractionated by ultrafiltration/size-exclusion chromatography (SEC) and eluted fractions were analysed by complementary techniques including immunoblotting, electron microscopy and nano-liquid chromatography-tandem mass spectrometry. A fraction of 90 K appeared as nanoparticles of irregular shape with heterogeneous dimensions of 15–60 nm that co-eluted with extracellular vesicles in SEC. Median levels of 90 K quantified by ELISA were not different between ALS patients (215.8 ng/ml) and controls (213.3 ng/ml) in contrast with the benchmark biomarker for ALS phosphoneurofilament heavy chain (1750 and 345 pg/ml, respectively). A multiregression model supported age is the only independent predictor of 90 K level in both groups (p < 0.05). Significant correlation was found between 90 K levels and age for the ALS group (r = 0.366, p = 0.031) and for all subjects (r = 0.392, p = 0.003). In conclusion, this study unveils the presence of 90 K-containing nanoparticles in human CSF and opens novel perspectives to further investigate 90 K as potential aging marker.
Highlights
Galectin-3 binding protein (LGALS3BP or 90 K) is a secreted glycoprotein found in human body fluids
cerebrospinal fluid (CSF) was fractionated by ultrafiltration/size-exclusion chromatography (SEC) and eluted fractions were analysed by complementary techniques including immunoblotting, electron microscopy and nano-liquid chromatography-tandem mass spectrometry
90 K was visualized by electron microscopy in nanoparticles of irregular shape found in extracellular vesicle (EV) fractions from tumor cells in culture[5]
Summary
Galectin-3 binding protein (LGALS3BP or 90 K) is a secreted glycoprotein found in human body fluids. A fraction of 90 K appeared as nanoparticles of irregular shape with heterogeneous dimensions of 15–60 nm that co-eluted with extracellular vesicles in SEC. Galectin-3-binding protein (LGALS3BP, known as Mac-2BP or tumor-associated antigen 90 K) is a secretory protein with 567 amino acid residues[1]. Purified full-length recombinant glycoprotein from human embryonic kidney cells was found to self-associate into non-covalent oligomers of 1000–1500 kDa that appeared as ring-like structures and other shapes by electron microscopy. 90 K was visualized by electron microscopy in nanoparticles of irregular shape found in extracellular vesicle (EV) fractions from tumor cells in culture[5]. 3 ml CSF were purified; for 90 K, IgG, albumin 10% of each eluted fraction were used, whereas for CD63, CD9 and L1CAM 30% were used
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