2N5R

NMR structure of cFLIP-derived calmodulin binding peptide

Summary for 2N5R

• Entry DOI: 10.2210/pdb2n5r/pdb
• NMR Information: BMRB: 25726
• Descriptor: CASP8 and FADD-like apoptosis regulator (1 entity in total)
• Functional Keywords: calmodulin, apoptosis
• Biological source: Homo sapiens (human)
• Total number of polymer chains: 1
• Total formula weight: 1561.98

Authors

Panaitiu, A.E. (deposition date: 2015-07-24, release date: 2015-11-18, Last modification date: 2024-05-15)

Primary citation

Gaidos, G.,Panaitiu, A.E.,Guo, B.,Pellegrini, M.,Mierke, D.F.
Identification and Characterization of the Interaction Site between cFLIPL and Calmodulin.
Plos One, 10:e0141692-e0141692, 2015

PubMed Abstract: Overexpression of the cellular FLICE-like inhibitory protein (cFLIP) has been reported in a number of tumor types. As an inactive procaspase-8 homologue, cFLIP is recruited to the intracellular assembly known as the Death Inducing Signaling Complex (DISC) where it inhibits apoptosis, leading to cancer cell proliferation. Here we characterize the molecular details of the interaction between cFLIPL and calmodulin, a ubiquitous calcium sensing protein. By expressing the individual domains of cFLIPL, we demonstrate that the interaction with calmodulin is mediated by the N-terminal death effector domain (DED1) of cFLIPL. Additionally, we mapped the interaction to a specific region of the C-terminus of DED1, referred to as DED1 R4. By designing DED1/DED2 chimeric constructs in which the homologous R4 regions of the two domains were swapped, calmodulin binding properties were transferred to DED2 and removed from DED1. Furthermore, we show that the isolated DED1 R4 peptide binds to calmodulin and solve the structure of the peptide-protein complex using NMR and computational refinement. Finally, we demonstrate an interaction between cFLIPL and calmodulin in cancer cell lysates. In summary, our data implicate calmodulin as a potential player in DISC-mediated apoptosis and provide evidence for a specific interaction with the DED1 of cFLIPL.

PubMed: 26529318
DOI: 10.1371/journal.pone.0141692

Experimental method

SOLUTION NMR