Differential display of peptides induced during the immune response of Drosophila: A matrix-assisted laser desorption ionization time-of-flight mass spectrometry study

Sandrine Uttenweiler-Joseph (1) , Marc Moniatte (2) , Marie Lagueux (1) , Alain van Dorsselaer (3) , Jules A. Hoffmann (4) , Philippe Bulet (4)

1 IBMC - Institut de biologie moléculaire et cellulaire
2 Proteomics Core Facility [Lausanne, Suisse]
3 LSMBO - Laboratoire de Spectrométrie de Masse BioOrganique [Strasbourg]
4 RIDI - UPR 9002 - Réponse immunitaire et developpement chez les insectes

We have developed an approach based on a differential mass spectrometric analysis to detect molecules induced during the immune response of Drosophila, regardless of their biological activities. For this, we have applied directly matrix-assisted laser desorption/ionization MS to hemolymph samples from individual f lies before and after an immune challenge. This method provided precise information on the molecular masses of immune-induced molecules and allowed the detection, in the molecular range of 1.5-11 kDa, of 24 Drosophila immune-induced molecules (DIMs). These molecules are all peptides, and four correspond to already characterized antimicrobial peptides. We have further analyzed the induction of the various peptides by immune challenge in wild-type f lies and in mutants with a compromised antimi-crobial response. We also describe a methodology combining matrix-assisted laser desorption ionization time-off light MS, HPLC, and Edman degradation, which yielded the peptide sequence of three of the DIMs. Finally, molecular cloning and Northern blot analyses revealed that one of the DIMs is produced as a prepropeptide and is inducible on a bacterial challenge.

Domaines: Sciences du Vivant [q-bio]

Sandrine Uttenweiler-Joseph, Marc Moniatte, Marie Lagueux, Alain van Dorsselaer, Jules A. Hoffmann, et al.. Differential display of peptides induced during the immune response of Drosophila: A matrix-assisted laser desorption ionization time-of-flight mass spectrometry study. Proceedings of the National Academy of Sciences of the United States of America, 1998, 95 (19), pp.11342 - 11347. ⟨10.1073/pnas.95.19.11342⟩. ⟨hal-02513060⟩