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Reproducible label-free quantitative proteomics profiling cellular responses to engineered nanomaterials

ABSTRACT: Proteomic data from engineered nanomaterial- treated THP-1 cells. Time points at 12 hours; 4 biological replicates for each treatment. Human THP-1 cells were either mock-treated (control) or treated with engineered nanomaterials (ENMs). Four biological replicates were used for each sample group (Control, or ENM treatment at either the low or high dosage). In the block design scheme for LC-MS analysis, samples treated with two different ENMs were assign into an analytical block (2 ENM X 2 dosages/ENM X 4 replicates = 16 samples). Four control samples were also included in each block, yielding 20 samples per block. Blocks of samples were divided into two processing batches. Samples in each batch were processed simultaneously from cell culture, to treatment with ENMs, and finally to proteomic sample preparation for LC-MS analysis (including trypsin digestion).

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Wei-Jun Qian

PROVIDER: MSV000084903 | MassIVE | Wed Feb 05 00:17:00 GMT 2020

SECONDARY ACCESSION(S): PXD017388

REPOSITORIES: MassIVE

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Project description:During the last decades, the use of plant growth promoting bacteria (PGPB) has been found to increase crop yield and quality and to confer abiotic and biotic stress tolerance. However, until now the PGPB mechanism to enhance plant performances is not clearly defined. Recently, our findings demonstrated that inoculations with both Kocuria rhizophila and Streptomyces violaceoruber, as well as their combination, determined an increase of tomato (Solanum lycopersicum) growth and development. In this study, through an advanced differential proteomic approach on tomato leaves, plant molecular mechanisms affected by both K. rhizophila and S. violaceoruber have been elucidated. To this aim, tomato plants were treated with K. rhizophila and/or Streptomyces violaceoruber cultures and grown on coconut fiber in greenhouse. In particular, PGPB treatments were conducted twice, on seed and after two weeks from the seedling by fertirrigation. Thus, the analyses have been performed at 14 days after sowing (DAS) (T1) and 42 DAS (T2). The results confirmed the growth stimulation ability of K. rhizophila/Streptomyces violaceoruber, showing shoot fresh and dry weight significantly improved at each time sampling. For the early phase (DAS-T1) comparative proteomics analysis of tomato plant leaves, 2 biological replicates were set up for the plants used as control (i.e. not subjected to treatment - samples I1 and I2-control I), 2 biological replicates for plants subjected to treatment with K. rhizophila (samples L1 and L2-treatment L), 2 biological replicates for plants subjected to treatment with S. violaceoruber (samples M1 and M2-treatment M), and 2 biological replicates for plants subjected to treatment with a mix of the two bacterial strains (samples N1 and N2-treatment N), for a total of 8 samples of leaf protein extracts. For the late phase (DAS-T2) comparative proteomics analysis of tomato plant leaves, 2 biological replicates were set up for the plants used as control (i.e. not treated - samples A1 and A2 - control A), 2 biological replicates for plants subjected to treatment with K. rhizophila (samples B1 and B2-treatment B), 2 biological replicates for plants subjected to treatment with S. violaceoruber (samples C1 and C2-treatment C), and 2 biological replicates for plants subjected to treatment with a mix of the two bacterial strains (samples D1 and D2-treatment D), for a total of 8 samples of leaf protein extracts. Proteomic analysis was able to identify 239 and 203 significantly differentially represented proteins (DRPs) at T1 and T2, respectively, comparing PGPB-treated vs. untreated control plants. KEGG Orthology (KO) identified DRP belonging to photosynthesis, biosynthesis of secondary metabolites, and carbon metabolism.

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Reproducible label-free quantitative proteomics profiling cellular responses to engineered nanomaterials

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