Glycosylation patterns of human alpha2-macroglobulin: analysis of lectin binding by electron microscopy.
Human alpha2-macroglobulin (alpha 2M) is a 720 kDa glycoprotein that presents two ultrastructural conformations: slow (S-alpha 2M) and fast (F-alpha 2M). alpha 2M acts mainly as a proteinase scavenger, but an immunomodulatory role was also proposed. This work studies the effect of desialylation and deglycosylation on the structure patterns of alpha 2M by ultrastructural analysis of lectin-induced aggregates, which represents a new approach that had never been previously used. Transmission electron microscopy (TEM) analysis showed the loss of S-alpha 2M conformation after deglycosylation, indicating that glycosidic side-chains contribute to the molecular stability of S-alpha 2M. TEM proved to be an important tool to analyze the effect of biochemical changes on alpha 2M, yielding an objective qualitative control of its morphological state. Certain carbohydrate residues did not vary between the alpha 2M conformations, since both bound similarly ConA and WGA lectins. However, the binding of PNA and BSI-B(4) was slightly lower in F-alpha 2M than in S-alpha 2M. Among the neuraminidases used to desialylate both conformations of alpha 2M that from Arthrobacter ureafaciens was the most effective. Incubation with the lectins ConA or SNA, respectively specific for mannosyl and sialyl residues, led to dose-dependent patterns of aggregation of alpha 2M molecules, mediated by lectin binding and clearly visualized by TEM.
Paiva
M M
MM
Lab. de Microscopia Eletrônica e Microanálise, INT-NE/CETENE - Centro de Tecnologias Estratégicas do Nordeste, Recife, PE, Brazil. mauricio@cetene.gov.br
Soeiro
M N C
MN
Barbosa
H S
HS
Meirelles
M N L
MN
Delain
E
E
Araújo-Jorge
T C
TC
D006001:Glycoconjugates / Q000032:analysis*; D006801:Humans; D037102:Lectins / Q000378:metabolism; D046529:Microscopy, Electron, Transmission / Q000379:methods; D011485:Protein Binding; D011487:Protein Conformation; D000511:alpha-Macroglobulins / Q000737:chemistry* / Q000648:ultrastructure
D006001:Glycoconjugates; D037102:Lectins; D000511:alpha-Macroglobulins