Glycoconjugate Journal

, 25:741

Antiproliferative effect of T/Tn specific Artocarpus lakoocha agglutinin (ALA) on human leukemic cells (Jurkat, U937, K562) and their imaging by QD-ALA nanoconjugate

  • Urmimala Chatterjee
  • Partha Pratim Bose
  • Sharmistha Dey
  • Tej P. Singh
  • Bishnu P. Chatterjee
Article

DOI: 10.1007/s10719-008-9134-8

Cite this article as:
Chatterjee, U., Bose, P.P., Dey, S. et al. Glycoconj J (2008) 25: 741. doi:10.1007/s10719-008-9134-8

Abstract

T/Tn specificity of Artocarpus lakoocha agglutinin (ALA), isolated from the seeds of A. lakoocha (Moraceae) fruit and a heterodimer (16 kD and 12 kD) of molecular mass 28 kD, was further confirmed by SPR analysis using T/Tn glycan containing mammalian glycoproteins. N-terminal amino acid sequence analysis of ALA showed homology at 15, 19–21, 24–27, and 29 residues with other lectin members of Moraceae family viz., Artocarpus integrifolia (jacalin) lectin, Artocarpus hirsuta lectin, and Maclura pomifera agglutinin. It is mitogenic to human PBMC and the maximum proliferation was observed at 1 ng/ml. It showed an antiproliferative effect on leukemic cells, with the highest effect toward Jurkat cells (IC50 13.15 ng/ml). Synthesized CdS quantum dot-ALA nanoconjugate was employed to detect the expression of T/Tn glycans on Jurkat, U937, and K562 leukemic cells surfaces as well as normal lymphocytes by fluorescence microscopy. No green fluorescence was observed with normal lymphocytes indicating that T/Tn determinants, which are recognized as human tumor associated structures were cryptic on normal lymphocyte surfaces, whereas intense green fluorescent dots appeared during imaging of leukemic cells, where such determinants were present in unmasked form. The above results indicated that QD-ALA nanoconjugate is an efficient fluorescent marker for identification of leukemic cell lines that gives rise to high quality images.

Keywords

ALAArtocarpus lakoocha agglutininSurface plasmon resonanceLeukemic cellsQuantum dotNanoconjugate

Abbreviations

ALA

Artocarpus lakoocha agglutinin

BSM

Bovine submandibular gland mucin

CdS

Cadmium sulphide

ELLSA

Enzyme - linked lectinsorbent assay

ESI-MS

Electron spray ionization mass spectrometry

FPLC

Fast protein liquid chromatography

HBS

Hepes buffered saline

HEPES

N-(2-hydoxyethyl) piperizine-N′-(2-hydroxypropane sulfonic acid)

HSM

Hamster submaxillary mucin

PBMC

Peripheral blood mononuclear cells

PVDF

Polyvinylidene difluoride

Q-ToF

Quadrupole-time of flight

QD

Quantum dot

SDS-PAGE

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

2 ME

2-Mercaptoethanol

Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Urmimala Chatterjee
    • 1
    • 4
  • Partha Pratim Bose
    • 1
    • 5
  • Sharmistha Dey
    • 2
  • Tej P. Singh
    • 2
  • Bishnu P. Chatterjee
    • 1
    • 3
  1. 1.Department of Biological ChemistryIndian Association for the Cultivation of ScienceJadavpurIndia
  2. 2.Department of BiophysicsAll India Institute of Medical SciencesAnsari NagarIndia
  3. 3.Department of BiotechnologyWest Bengal University of TechnologyKolkataIndia
  4. 4.Department of Anatomy, Physiology and BiochemistrySwedish University of Agricultural ScienceUppsalaSweden
  5. 5.Department of Biochemistry and Organic ChemistryUppsala UniversityUppsalaSweden