Mannose-specific lectins bind alpha-2-macroglobulin and an unknown protein from human plasma.
GNA, the mannose-specific lectin from Galanthus nivalis was confirmed to bind alpha-2-macroglobulin (A2M) but another protein was copurified with A2M from total human plasma. A total of 23 other lectins with diverse specificities were tested for reaction with human A2M and with three other members of the A2M family. NPA, a mannose-specific lectin isolated from Narcissus pseudonarcissus bulbs, and RSA, the Rhizoctonia solani agglutinin, were selected for further testing. For isolation of A2M, immobilized NPA was superior to GNA because its binding capacity was an order of magnitude higher. The specificity of these lectins must be very similar however, because the same unknown plasma protein was also bound by NPA. A2M and the unknown protein must share a unique mannose carbohydrate structure not present in any other human plasma protein. The copurified protein subunit size of 185 kDa is very similar to that of A2M, but the native molecular mass of 350 kDa indicated a noncovalent homodimer structure. Together with the acid isoelectric point this is not typical for any known plasma protein nor for any unidentified spot on the two-dimensional map of human plasma proteins. No immunological reaction with available antisera was evident. A specific antiserum raised to the unknown protein demonstrated its presence in all human plasma samples examined. The N-terminal residue was blocked, whereas internal protein sequences obtained after CNBr fragmentation and proteolysis were not homologous to any known protein sequence. These data demonstrate that this protein is unknown and not a proteinase inhibitor of the A2M family.
Van Leuven
F
F
Center of Human Genetics, Katholieke Universiteit van Leuven, Belgium.
Torrekens
S
S
Van Damme
E
E
Peumans
W
W
Van den Berghe
H
H
D000595:Amino Acid Sequence; D001798:Blood Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods*; D027801:Galanthus; D006801:Humans; D037102:Lectins / Q000378:metabolism*; D008358:Mannose / Q000378:metabolism*; D037241:Mannose-Binding Lectins; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000235:genetics; D037121:Plant Lectins; D010949:Plasma / Q000737:chemistry*; D012680:Sensitivity and Specificity; D000511:alpha-Macroglobulins / Q000302:isolation & purification* / Q000378:metabolism
D001798:Blood Proteins; D037102:Lectins; D037241:Mannose-Binding Lectins; D010446:Peptide Fragments; D037121:Plant Lectins; D000511:alpha-Macroglobulins; C063117:snowdrop lectin; D008358:Mannose