A neutrophil migration-inducing lectin from Artocarpus integrifolia.

Abstract

A neutrophil migration-inducing protein has been isolated from the saline extract of Artocarpus integrifolia seeds by successive sugar affinity chromatography steps during which the protein was not absorbed by D-galactose resin, and then was absorbed to and eluted from D-mannose resin by 0.1 M D-mannose. Gel filtration on Superdex 75 HR indicated a molecular mass of 52 kDa when 0.1 M D-mannose was present in the elution buffer. A single band of apparent molecular mass of 13 kDa was demonstrable by SDS-PAGE only after heating, both in the presence and absence of reducing agent, suggesting that the molecule is a tetramer formed by the noncovalent association of 13 kDa chains. Isoelectric forms corresponding to isoelectric points of 4.0, 4.2, 5.0, and 5.2 were demonstrable by isoelectric focusing-PAGE, and four active forms having the same isoelectric points were separated by chromatofocusing. The minimal m.w. calculated from amino acid analysis data was 13,193. The protein, denoted KM+, stimulated neutrophil migration in the rat peritoneal cavity assay in a dose-related manner in the range of 1 to 300 micrograms per rat. The dose-response curve of the in vitro chemotactic activity of KM+ was bell shaped and its ascending limb was dose dependent in the range of 1 ng to 10 micrograms/well. D-Mannose (0.1 M) inhibited the in vitro (80%) and in vivo (60%) neutrophil migration-inducing activities of KM+ and also its hemmaglutinating activity. The chemotactic activity was shown to be caused by haptotaxis rather than chemokinesis. The physical and biologic properties of KM+ suggest that this lectin may attract neutrophils by a mechanism involving a haptotactic gradient as has been proposed for IL-8. KM+ might be used as tool to study protein-carbohydrate interactions during neutrophil migration through the extracellular matrix.

ArtinM Cytotoxicity in B Cells Derived from Non-Hodgkin’s Lymphoma Depends on Syk and Src Family Kinases

“…Our group previously reported the immunomodulatory activity of ArtinM, a D-mannose-binding lectin obtained from the seeds of Artocarpus heterophyllus . ArtinM is structurally organized as a homotetramer formed by 16-kDa non-glycosylated subunits, each polypeptide chain containing a CRD with an affinity for Manα1–3 [Manα1–6] Manβ1–4, which constitutes the N-glycans core [ 19 , 20 , 21 ]. Immunomodulation of the Th1 phenotype by ArtinM occurs via its interaction with TLR2/CD14 N-glycans on antigen-presenting cells, which induces IL-12 production [ 8 , 22 ].…”

Estudo da atividade inseticida e pro-inflamatoria da lectina isolada de sementes de 'Annona coriacea' Mart

“…Algumas lectinas de plantas podem mimetizar as lectinas endógenas de mamíferos e induzir a migração de neutrófilos e células mononucleares in vivo e in vitro (Bento et al, 1993;Santos de Oliveira et al, 1994). Mais recentemente, foi demonstrado que a lectina de soja também pode induzir a migração de neutrófilos e de linfócitos in vivo e ativar as células mononucleares (Benjamim et al, 1997).…”

“…O carboidrato D-mannose, após incubação com a lectina, acentuadamente reduziu a migração de neutrófilos induzida por ACLEC. Estes dados são semenlhantes aos encontrados em estudos anteriores que demonstram que este mesmo carboidrato foi capaz de inibir a migração de neutrófilos induzida por lectinas purificadas de sementes de Artocarpus integrifolia (Santos de Oliveira et al, 1994), Talisia esculenta (Freire et al, 2003) e Pisum arvense (Alencar et al, 2005). Outros carboidratos tais como glicose e galactose inibiram o influxo de neutrófilos na cavidade peritoneal de ratos em resposta a lectinas de Canavalia brasiliensis, Dioclea grandiflora (Bento et al, 1993), Glycine max (Benjamin et al, 1997) e Vatairea macrocarpa (Alencar, et al, 2003).…”