Figure 1 | Scientific Reports

Figure 1

From: Calpain cleavage of Junctophilin-2 generates a spectrum of calcium-dependent cleavage products and DNA-rich NT1-fragment domains in cardiomyocytes

Figure 1

Identification of Calpain-specific cleavage fragments of JP2. (A) Mouse FL JP2 domain representation (Uniprot: Q9ET78) summarizing the in silico prediction of Calpain cleavage sites by the DeepCalpain tool. Predicted cleavage sites are indicated by vertical black lines above the JP2 topology domain model. Experimentally confirmed cleavage events observed at 0.1 U/ml Calpain-2 are indicated by scissors. The approximate epitope binding positions of three different JP2 antibodies are indicated by Y-shaped symbols: NT, N-terminal region; M, middle region; CT, C-terminal region. (B) The Calpain-2 activity specific cleavage patterns of the mouse FL JP2 substrate. Purified recombinant JP2 was incubated at increasing Calpain-2 concentrations for 30 min at 30 °C, followed by SDS-PAGE and Coomassie staining. (C) Box plot summarizing the JP2 cleavage events (scissors), fragments, and cleavage sites experimentally observed for the 0.1 U/ml Calpain-2 concentration. (D) LC-MS/MS analysis of the spectrum of mouse JP2 fragments generated by Calpain-2 digestion. Tryptic peptide counts are represented as relative abundance versus the JP2 amino acid position. Arrows indicate the positions of the first, second, and third cleavage events.

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