title: Antibody-mediated sialidase activity in blood serum of patients with multiple myeloma.
abstract: Cell surface sialylation is known to be tightly connected with tumorigenicity, invasiveness, metastatic potential, clearance of aged cells, while the sialylation of IgG molecules determines their anti-inflammatory properties. Four sialidases - hydrolytic enzymes responsible for cleavage of sialic residues - were described in different cellular compartments. However, sialidases activity in body fluids, and specifically in blood serum, remains poorly studied. Here, we characterize first known IgG antibodies possessing sialidase-like activity in blood serum of multiple myeloma (MM) patients. Ig fractions were precipitated with ammonium sulfate (50% of saturation) from blood serum of 12 healthy donors and 14 MM patients, and screened for the presence of sialidase activity by using 4-MUNA (2'-(4-methylumbelliferyl)-α-D-N-acetylneuraminic acid) as substrate. High level of sialidase activity was detected in the MM patients, but not in healthy donors. Subsequent antibody purification by protein-G affinity chromatography and HPLC size exclusion chromatography at acidic conditions demonstrated that sialidase activity was attributable to IgG molecules. Sialidase activity was also specific for (Fab)(2) fragment of IgG and blocked by sialidase inhibitor DANA. Sialidase activity of IgG molecule was also confirmed by in gel assay for cleavage of sialidase substrate. Kinetic parameters of the catalysis reaction were described by Michaelis-Menten equation with K(m)  = 44.4-108 µM and k(cat) = 2.7-23.1 min(-1). The action of IgG possessing sialidase-like activity towards human red blood cells resulted in a subsequent increase in their agglutination by the peanut agglutinin, that confirms their desialylation by the studied IgG. This is the first demonstration of the intrinsic sialidase activity of IgG isolated from blood serum of MM patients.
authors.affiliation: Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv 79005, Ukraine.
authors.forename: Rostyslav
authors.initials: R
authors.lastname: Bilyy
authors.forename: Andriy
authors.initials: A
authors.lastname: Tomin
authors.forename: Iryna
authors.initials: I
authors.lastname: Mahorivska
authors.forename: Olga
authors.initials: O
authors.lastname: Shalay
authors.forename: Volodymyr
authors.initials: V
authors.lastname: Lohinskyy
authors.forename: Rostyslav
authors.initials: R
authors.lastname: Stoika
authors.forename: Yuriy
authors.initials: Y
authors.lastname: Kit
mesh_terms: D000328:Adult; D017151:Antibodies, Catalytic / Q000378:metabolism*; D015153:Blotting, Western; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009101:Multiple Myeloma / Q000201:enzymology*; D009439:Neuraminidase / Q000097:blood / Q000378:metabolism*
chemical_list: D017151:Antibodies, Catalytic; D009439:Neuraminidase