Abstract
In this work, we compared the proteomic profiles of outer membrane vesicles (OMVs) isolated from Rhizobium etli CE3 grown in minimal medium (MM) with and without exogenous naringenin. One-hundred and seven proteins were present only in OMVs from naringenin-containing cultures (N-OMVs), 57 proteins were unique to OMVs from control cultures lacking naringenin (C-OMVs) and 303 proteins were present in OMVs from both culture conditions (S-OMVs). Although we found no absolute predominance of specific types of proteins in the N-, C- or S-OMV classes, there were categories of proteins that were significantly less or more common in the different OMV categories. Proteins for energy production, translation and membrane and cell wall biogenesis were overrepresented in C-OMVs relative to N-OMVs. Proteins for carbohydrate metabolism and transport and those classified as either general function prediction only, function unknown, or without functional prediction were more common in N-OMVs than C-OMVs. This indicates that naringenin increased the proportion of these proteins in the OMVs, although NodD binding sites were only slightly more common in the promoters of genes for proteins found in the N-OMVs. In addition, OMVs from naringenin-containing cultures contained nodulation factor.
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Acknowledgements
We thank Dr. Carmen Quinto (Instituto de Biotecnología-UNAM) for providing R. etli strain UBP102. Part of this work was supported by CONACyT Grant 220790 and DGAPA-PAPIIT Grants IN213216 and IN207519.
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Fig. S1.
Maldi-TOF mass spectrometry analysis of n-butanol extracts of OMVs and supernatant from R. etli CE3 and a CE3 nodA− mutant. The strains were grown in of two liters of MM inoculated to an initial optical density of 0.5 at 540 nm OMVs were purified as described in Methods, extracted with n-butanol and the extract dried by a lyophilization. The dried residue was resuspended in distilled sterile water and analyzed by HPLC on a reverse phase C-18 column (Nova pack C18 3.9x150 mm Waters) with detection at 215 nm using CH3CN 20% as mobile phase. Peak fractions were collected lyophilized, and dissolved in methanol/water (1:1). Five µl samples were injected in the masses spectrophotometer, run was done in a LTQ XL-Orbitrap mass spectrometer (Thermo Fisher Scientific, Bremen, Germany) in a positive mode by DIC. Ion masse data were analyzed using the Glycomod (htpp://www.expasy.org/tools/glycomod_masses.html) data base. The ion m/z peak of 1500.9 for the Nod factor coincided with that reported previously (Cárdenas et al. 1995). (PDF 77 kb)
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Taboada, H., Dunn, M.F., Meneses, N. et al. Qualitative changes in proteins contained in outer membrane vesicles produced by Rhizobium etli grown in the presence of the nod gene inducer naringenin. Arch Microbiol 201, 1173–1194 (2019). https://doi.org/10.1007/s00203-019-01682-4
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DOI: https://doi.org/10.1007/s00203-019-01682-4