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A neutrophil migration-inducing lectin from Artocarpus integrifolia.

R Santos-de-Oliveira, M Dias-Baruffi, S M Thomaz, L M Beltramini and M C Roque-Barreira
J Immunol August 15, 1994, 153 (4) 1798-1807;
R Santos-de-Oliveira
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M Dias-Baruffi
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S M Thomaz
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L M Beltramini
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M C Roque-Barreira
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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.

  • Copyright © 1994 by American Association of Immunologists

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The Journal of Immunology
Vol. 153, Issue 4
15 Aug 1994
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A neutrophil migration-inducing lectin from Artocarpus integrifolia.
R Santos-de-Oliveira, M Dias-Baruffi, S M Thomaz, L M Beltramini, M C Roque-Barreira
The Journal of Immunology August 15, 1994, 153 (4) 1798-1807;

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A neutrophil migration-inducing lectin from Artocarpus integrifolia.
R Santos-de-Oliveira, M Dias-Baruffi, S M Thomaz, L M Beltramini, M C Roque-Barreira
The Journal of Immunology August 15, 1994, 153 (4) 1798-1807;
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Print ISSN 0022-1767        Online ISSN 1550-6606