Journal: Molecular Cell

Article Title: Evolutionary origins and interactomes of human, young microproteins and small peptides translated from short open reading frames

doi: 10.1016/j.molcel.2023.01.023

Figure Lengend Snippet: Peptide interactomes can predict modulators of cellular functions (A) GO enrichment analysis of all interacting proteins of 16 ribosome-binding peptides 3–15 aa compared with all other peptides. (B) Violin plot with hydrophobicity values of the 16 ribosome-binding peptides 3–15 aa compared with all other peptides 3–15 aa . Horizontal lines indicate the mean ± standard deviation. (C) Number of arginines of the 16 ribosome-binding peptides 3–15 aa compared with all other peptides 3–15 aa , normalized to the total number of amino acids. (D) Schematic and results of the luciferase reporter assay performed with five randomly selected ribosome-binding peptides 3–15 aa . The significance was calculated using ANOVA and Tukey post hoc test. (E) Volcano plots of four AP-binding peptides 3–15 aa . Proteins assigned to the GO term vesicle-related transport (GO:0016192) are highlighted in red. (F) Circos plot of all peptides 3–15 aa that interact with endocytic proteins. (G) Peptide sequences of the four AP-binding peptides 3–15 aa (aromatic aa highlighted in red, di-hydrophobic motifs underlined) and GO enrichment analysis of their interactomes. (H) Representative immunofluorescence images of fluorescently labeled RAP internalized by BN16 cells treated with DMSO, dynasore, PPARD- and ARMC1-uORF-peptide, respectively. Scale bar represents 200 μm. (I) Results of the RAP endocytosis assay (five replicates per condition). Values were normalized to total protein content, and samples without RAP treatment were subtracted and then normalized to the treatment with RAP only (=100%). The PPARD-uORF-peptide, which did not bind APs, was included as a control ( Figure S6 J). The statistical significance was calculated using ANOVA and Tukey post hoc test.

Article Snippet: Brown Norway rat yolk sac carcinoma (BN16) cells (CRL-2180, ATCC, sex unknown) were cultivated in DMEM supplemented with 10% FBS and 1% Penicillin/Streptomycin.

Techniques: Binding Assay, Standard Deviation, Luciferase, Reporter Assay, Immunofluorescence, Labeling, Endocytosis Assay, Control

Journal: Molecular Cell

Article Title: Evolutionary origins and interactomes of human, young microproteins and small peptides translated from short open reading frames

doi: 10.1016/j.molcel.2023.01.023

Figure Lengend Snippet:

Article Snippet: Brown Norway rat yolk sac carcinoma (BN16) cells (CRL-2180, ATCC, sex unknown) were cultivated in DMEM supplemented with 10% FBS and 1% Penicillin/Streptomycin.

Techniques: Transduction, Recombinant, Membrane, Protease Inhibitor, In Situ, Proximity Ligation Assay, Labeling, Luciferase, Phospho-proteomics, Knock-Out, RNA Sequencing, Sequencing, Microscopy, Immunofluorescence, Software, Targeted Proteomics