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Galectins pp 169-184 | Cite as

Examination of Galectin-Induced Lattice Formation on Early B-Cell Development

  • Stéphane J. C. Mancini
  • Latifa Elantak
  • Annie Boned
  • Marion Espéli
  • Françoise Guerlesquin
  • Claudine SchiffEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1207)

Abstract

Galectin-1 (GAL1) is a pre-B cell receptor (pre-BCR) ligand that induces pre-BCR clustering and leads to efficient pre-B cell proliferation and differentiation in the bone marrow. To study pre-BCR–GAL1 interactions and its functional consequence on the early steps of the B cell development, we combine structural nuclear magnetic resonance (NMR) approaches and B cell biology techniques. NMR is applied to identify the residues involved in pre-BCR–GAL1 interactions by monitoring chemical shift perturbations when the complex is formed. This structural information is then used at the cellular level to target specifically the complex formation during GAL1-induced pre-BCR clustering and lattice formation, using immunofluorescence techniques. Moreover, an in vivo assay was set up to study the consequence of synapse formation on the early steps of B cell development.

Key words

Galectin-1 pre-B cell receptor Lattice formation Protein interactions B cell development Nuclear magnetic resonance Immunofluorescence Flow cytometry 

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Stéphane J. C. Mancini
    • 1
  • Latifa Elantak
    • 2
  • Annie Boned
    • 1
  • Marion Espéli
    • 3
  • Françoise Guerlesquin
    • 2
  • Claudine Schiff
    • 1
    Email author
  1. 1.Centre d’Immunologie de Marseille-Luminy, INSERM U1104, CNRS UMR7280, case 906Aix-Marseille Université, UM2Marseille cedex 09France
  2. 2.Laboratoire d’Ingénierie des Systèmes Macromoléculaires, CNRS UMR7255CNRS, Aix-Marseille UniversitéMarseilleFrance
  3. 3.University of Cambridge School of Clinical MedicineCambridgeUK

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