Lysosomes are the main degradative organelles of cells and involved in a variety of processes including the recycling of macromolecules, storage of compounds, and metabolic signaling. Despite an increasing interest in the proteomic analysis of lysosomes, no systematic study of sample preparation protocols for lysosome enriched fractions has been performed to date. In the current study, we used samples enriched for lysosomes by paramagnetic nanoparticles and systematically evaluated experimental parameters for the analysis of the lysosomal proteome. This includes different approaches for the concentration of lysosome-containing fractions; desalting of samples by solid phase extraction; fractionation of peptide samples; and different gradient lengths for LC-MS/MS analyses of unfractionated samples by data dependent and data independent acquisition. Furthermore, we evaluated four different digestion methods including filter aided sample preparation (FASP), in-gel digestion, and in-solution digestion using either RapiGest or urea. Using the combined data, we generated a benchmark lysosomal proteome data set for mouse embryonic fibroblasts as well as a spectral library for the analysis of lysosomes by data independent acquisition.

No abstract

An extract of nishyinda (Vitex negundo) leaves, black pepper (Piper nigrum) and cinnamon (Cinnamomum verum) (polyherbal extract) were used as growth promoter in broilers. A total of 20 day-old broiler chicks were purchased and after seven days of acclimatization randomly divided into two equal groups. No vaccination schedule was practised and no antibiotics were added in rations. Group A served as control while group B was supplemented with polyherbal extract 1 mL/litre in drinking water. Weekly body weight gain up to six weeks was measured and blood tests were performed at 21 and 42 days. Polyherbal extract significantly (P<0.05) improved weight gain. There was no change in haematological parameters. It can be concluded that the polyherbal extract was safe as a growth promoter in broiler production without adverse effects on chicken health. (Bangl.

Lysosomes play a key role in the regulation of cellular metabolism and are increasingly recognized as highly active and diverse organelles which are involved in a large variety of processes. Their essential role is exemplified by the detrimental consequences of lysosomal malfunction, which can result in lysosomal storage disorders, neurodegenerative diseases, and cancer. Using lysosome enrichment and mass spectrometry, we investigated the lysosomal proteomes of six common cell lines. We provide first evidence for cell-type specific differences of lysosomes on a large scale, showing highly variable levels of distinct lysosomal proteins within one cell type, while others are highly conserved among cell lines. Using stable isotope labelling and bimodal distribution analysis, we identify high confidence lysosomal proteins for each cell line. Multi cell line correlation of these data reveals potential novel lysosomal proteins, and we confirm lysosomal localization for five candidates. All data are available via ProteomeXchange with identifier PXD020600.