vflip-ikk g blocker
DESCRIPTION
vFLIP-IKK g Blocker . Edith Chan WIBR. IKK g. NF- k B Pathway. The Nf- k B pathway is related to inflammatory responses, cell death, and oncogenesis. Kaposi’s sarcoma herpesvirus (KSHV) encoded FLIP (vFLIP) binds to IKK g to activate NF k B. - PowerPoint PPT PresentationTRANSCRIPT
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vFLIP-IKK Blocker
Edith Chan
WIBR
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NF-B Pathway• The Nf-B pathway is related to
inflammatory responses, cell death, and oncogenesis.
• Kaposi’s sarcoma herpesvirus (KSHV) encoded FLIP (vFLIP) binds to IKK to activate NFB.
• Similarly, in human, cFLIP can associated with IKK and thus has a wider application in cancer therapies.
IKK
3
Work Study
• The study focuses on using X-ray crystal structures, biophysical screening and structure based design to identify blockers of the vFLIP-IKK and p22-cFLIP-IKK interaction, conducting lead optimisation and identifying a development candidate.
• My immediate actions are
– Understanding of the interaction of the vFLIP-IKKcomplex using X-ray crystal structure
– Selection of compounds mimicking the IKK interaction of the complex
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X-ray crystal structure• Our collaborators at BBK have
solved the structure between vFLIP-IKK (3cl3).
• Full length IKK is 419aa long mulitdomain protein
• Both proteins are truncated– ks-vFLIP (aa1-178) [188aa]– IKKg (aa150-272) [419aa]
• The X-ray structure comprised of a dimer of two ks-vFLIP-IKKcomplex.
• The two vFLIP molecules come together solely through interactions between the two IKK chains.
IKK
vFLIPC-terminus
N-terminus
A B
D
E
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Protein-Protein Interactions
• Each of the IKK helix is interacting with a copy of the vFLIP via two adjacent vertical clefts (cleft 1 and 2)
• Cleft 1 involved more interactions between the complex, the hottest spot seems to be around Phe238 (of IKK)
6193QQHSVQVDQLRMQGQSVEAALRMERQAASEEKRKLAQLQVAYHQLFQEYDNHIKSSVVGSE253
226 246
Peptide tool compound• Full length IKK is 419aa long mulitdomain protein. In
the X-ray structure, only the HLX2 (helix) domains (aa193-253) are co-crystallized with ks-vFLIP.
• As seen from the picture, only a portion of the IKK is interacting with vFLIP. They are about 20aa long as highlighted in a pink box.
• Our first step is to use of peptide as proof of concept tool in further experiments
• Can a shorter peptide of IKK be sufficient to bind with vFLIP?
• Can a shorter peptide adopt helical conformation on its own or by recognition to vFLIP?
IKK
vFLIP
N-terminus
C-terminus
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Proposed tool Peptides
Length AA spin1. Longer peptide include C-terminus 30aa aa224-253
2. Spin the entire interaction with vFLIP 21aa aa226-246
3. Peptide that covers cleft1 interaction 16aa aa231-246
4. Peptide that covers cleft2 interaction 15aa aa226-240
5. Peptide that covers all essential interaction 12aa aa231-242
193QQHSVQVDQLRMQGQSVEAALRMERQAASEEKRKLAQLQVAYHQLFQEYDNHIKSSVVGSE2531 QQHSVQVDQLRMQGQSVEAALRMERQAASEEKRKLAQLQVAYHQLFQEYDNHIKSSVVGSE2 QQHSVQVDQLRMQGQSVEAALRMERQAASEEKRKLAQLQVAYHQLFQEYDNHIKSSVVGSE3 QQHSVQVDQLRMQGQSVEAALRMERQAASEEKRKLAQLQVAYHQLFQEYDNHIKSSVVGSE4 QQHSVQVDQLRMQGQSVEAALRMERQAASEEKRKLAQLQVAYHQLFQEYDNHIKSSVVGSE 5 QQHSVQVDQLRMQGQSVEAALRMERQAASEEKRKLAQLQVAYHQLFQEYDNHIKSSVVGSE
226 238 246