Journal: PLoS ONE

Article Title: High-Resolution Analysis and Functional Mapping of Cleavage Sites and Substrate Proteins of Furin in the Human Proteome

doi: 10.1371/journal.pone.0054290

Figure Lengend Snippet: A flowchart representing the stepwise approach used to identify furin substrates in the human proteome.

Article Snippet: Where indicated in our figures and figure legends, cells were incubated for 16–18 h with a furin inhibitor decanoyl-Arg-Val-Lys-Arg-chloromethylketone (50 μM; DEC; Bachem).

Techniques:

Journal: PLoS ONE

Article Title: High-Resolution Analysis and Functional Mapping of Cleavage Sites and Substrate Proteins of Furin in the Human Proteome

doi: 10.1371/journal.pone.0054290

Figure Lengend Snippet: A . The structure of furin with the SSNSRKRR↓S modeled substrate. The structure of furin-DEC complex (PDB 1P8J) was used as a template for modeling the protein substrate (shown as sticks) in the furin structure. The molecular surface of furin is colored according to the electrostatic potential (red, blue and white are negative, positive and neutral electrostatic potential values, respectively). Close-up on the left shows the S6 Glu230 and S5 Glu257 (yellow), which are critical for the interactions with the P6 and P5 substrate sub-sites. Close-ups ( B and C ) show in a more detail the P5-P4 and P2-P1’ residue positions in the furin active site.

Article Snippet: Where indicated in our figures and figure legends, cells were incubated for 16–18 h with a furin inhibitor decanoyl-Arg-Val-Lys-Arg-chloromethylketone (50 μM; DEC; Bachem).

Techniques: Residue

Journal: PLoS ONE

Article Title: High-Resolution Analysis and Functional Mapping of Cleavage Sites and Substrate Proteins of Furin in the Human Proteome

doi: 10.1371/journal.pone.0054290

Figure Lengend Snippet: Counts of probable protein substrates of furin and cleavage sites determined by our computational-experimental approach.

Article Snippet: Where indicated in our figures and figure legends, cells were incubated for 16–18 h with a furin inhibitor decanoyl-Arg-Val-Lys-Arg-chloromethylketone (50 μM; DEC; Bachem).

Techniques:

Journal: PLoS ONE

Article Title: High-Resolution Analysis and Functional Mapping of Cleavage Sites and Substrate Proteins of Furin in the Human Proteome

doi: 10.1371/journal.pone.0054290

Figure Lengend Snippet: Classification of the predicted furin protein substrates (based on the 598 GI protein sequences).

Article Snippet: Where indicated in our figures and figure legends, cells were incubated for 16–18 h with a furin inhibitor decanoyl-Arg-Val-Lys-Arg-chloromethylketone (50 μM; DEC; Bachem).

Techniques:

Journal: PLoS ONE

Article Title: High-Resolution Analysis and Functional Mapping of Cleavage Sites and Substrate Proteins of Furin in the Human Proteome

doi: 10.1371/journal.pone.0054290

Figure Lengend Snippet: The original glioma U251 cells, breast carcinoma MCF-7 cells with the enforced expression of the β3 integrin subunit (MCF7-β3), colon carcinoma LoVo cells with and without the enforced expression of furin and CHO cells with the enforced expression of the integrin αV precursor (pro-αV) or the furin-resistant pro-αV mutant (pro-αV-A889A890) were cell-surface biotinylated and then lysed. The lysates were precipitated using the αVβ3 antibody LM609 and analyzed by Western blotting with Extravidin-horseradish peroxidase. Where indicated, cells were co-incubated for 16–18 h prior to biotinylation with a synthetic furin inhibitor (DEC; 50 μM).

Article Snippet: Where indicated in our figures and figure legends, cells were incubated for 16–18 h with a furin inhibitor decanoyl-Arg-Val-Lys-Arg-chloromethylketone (50 μM; DEC; Bachem).

Techniques: Expressing, Mutagenesis, Western Blot, Incubation

Journal: PLoS ONE

Article Title: High-Resolution Analysis and Functional Mapping of Cleavage Sites and Substrate Proteins of Furin in the Human Proteome

doi: 10.1371/journal.pone.0054290

Figure Lengend Snippet: Ingenuity Pathway Analysis was used to identify the most significant developmental functions ( A ), molecular and cellular functions ( B ) and canonical pathways ( C ) associated with the predicted furin protein substrates. Right-tailed Fisher exact test was used to calculate a p-value. Only pathways with P<0.05 are shown.

Article Snippet: Where indicated in our figures and figure legends, cells were incubated for 16–18 h with a furin inhibitor decanoyl-Arg-Val-Lys-Arg-chloromethylketone (50 μM; DEC; Bachem).

Techniques:

Journal: PLoS ONE

Article Title: High-Resolution Analysis and Functional Mapping of Cleavage Sites and Substrate Proteins of Furin in the Human Proteome

doi: 10.1371/journal.pone.0054290

Figure Lengend Snippet: A . Pluripotency in human embryonic stem cells; B . the bone morphogenic protein (BMP) pathway in cardiomyocyte differentiation; C . Notch signaling pathway. Furin substrates are pink.

Article Snippet: Where indicated in our figures and figure legends, cells were incubated for 16–18 h with a furin inhibitor decanoyl-Arg-Val-Lys-Arg-chloromethylketone (50 μM; DEC; Bachem).

Techniques: