IQUIBICEN   23947
INSTITUTO DE QUIMICA BIOLOGICA DE LA FACULTAD DE CIENCIAS EXACTAS Y NATURALES
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Identification of the human receptor for the Sthapylococcus aureus lipoprotein Lpl and its role in cellular invasion
Autor/es:
LUQMAN, ARIF; SAAS P; PAULA M. TRIBELLI; GOTZ F
Reunión:
Congreso; 8th Congress of European Microbiologists; 2019
Institución organizadora:
FEMS
Resumen:
Background: Staphylococcus aureus is an opportunistic human pathogenic bacterial species that causes serious infections. Lipoproteins encoded in the lpl cluster triggers the invasion of S. aureus in nonprofessional phagocytic cells (NPPCs). S. aureus internalization by NPPCs is based on the adhesion to the host cell resulting in signal transduction and internalization of the bacteria. However, the receptor for Lpl protein in the host cells is still unknown.Objectives: our objective is to determinate the receptor of Lpl1 in NPPCs.Methods: Lpl1 protein was used as prototype of the Lpl proteins. We used soluble Lpl1 lacking the lipid signal peptide and a his-tag (Lpl1-his(-sp) to facilitate its purification from the cytosol of S. aureus using standard methods. Pull-down experiments were performed using Lpl1-his(-sp) as bait bound to Ni+2-column; HaCat cell lysate was used as a source of the receptor. Elution fractions were analyzed by SDS and stained using Coomasie blue. Visible bands in the experimental samples and absent in the control (without Lpl1) were cut and analyzed by Mass Spectrometry (MS).Results: We successfully purified Lpl1-his(-sp) from S.aureus. In pull-down experiments we obtained three bands in which Lpl1-his(-sp) was interacting with a potential host cell receptor. The host proteins were analysed by MS analysis. We identified and discarded well-known cytosolic proteins regarding transcription, translation or metabolic functions. We selected two candidates and our analysis revealed that both proteins belong to a human chaperon family. The proteins are now under investigation.