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Figure 4 | BMC Microbiology

Figure 4

From: A novel receptor – ligand pathway for entry of Francisella tularensis in monocyte-like THP-1 cells: interaction between surface nucleolin and bacterial elongation factor Tu

Figure 4

Identification of bacterial ligands for nucleolin. Part A. Samples were prepared as outlined in Experimental Procedures, normalized to 50 μg proteins per lane before separation by SDS-PAGE. Immunoblotting (I.B.) was performed either with anti-EF-Tu Ab diluted 1/100,000 (part A1) or anti-IglA Ab diluted 1/5,000 (part A2). Part B. 500 μg LVS membrane proteins were incubated with different concentrations of biotinylated p63 peptide (p63*). Complexes, formed between bacteria proteins and biotinylated p63 peptide, were isolated by purification using avidin-agarose. Purified proteins were analyzed by SDS-PAGE. Immunoblotting (I.B.) was performed with anti-EF-Tu Ab diluted 1/100,000. Mobility of marker proteins is indicated on left side. Part C. Specific binding of EF-Tu present in LVS membranes with RGG domain of nucleolin. 500 μg LVS membrane proteins were preincubated without (lane -) or with 50 μM unlabeled p63 peptide, before incubation with 5 μM biotinylated p63 peptide (p63*). Complex formed between EF-Tu present in bacteria membrane proteins and biotinylated p63 peptide was analyzed by immunoblotting with anti-EF-Tu Ab diluted 1/100,000. In control, 50 μg LVS membrane proteins were run directly.

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BMC Microbiology

ISSN: 1471-2180

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