Sequence specific peptidomimetic molecules inhibitors of a protein-protein interaction at the helix 1 level of c-myc (Articolo in rivista)

Type
Label
  • Sequence specific peptidomimetic molecules inhibitors of a protein-protein interaction at the helix 1 level of c-myc (Articolo in rivista) (literal)
Anno
  • 2005-01-01T00:00:00+01:00 (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#doi
  • 10.1096/fj.04-2369fje (literal)
Alternative label
  • Nieddu E.; Melchiori A.; Pescarolo M.P.; Avignolo C.; Bagnasco L.; Balbi C.; Barboro P.; Biasotti B.; Carnemolla B.; Castagnino N.; Cimoli G.; Damonte G.; D'Arrigo C.; Fassina G.; Licheri B.; Malacarne D.; Orecchia P.; Palomba D.; Pasa S.; Patrone E.; Ponassi R;Scapolla A.; Tortolina L.; Vasile F.; Mazzei M.; Parodi S. (2005)
    Sequence specific peptidomimetic molecules inhibitors of a protein-protein interaction at the helix 1 level of c-myc
    in The FASEB journal
    (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#autori
  • Nieddu E.; Melchiori A.; Pescarolo M.P.; Avignolo C.; Bagnasco L.; Balbi C.; Barboro P.; Biasotti B.; Carnemolla B.; Castagnino N.; Cimoli G.; Damonte G.; D'Arrigo C.; Fassina G.; Licheri B.; Malacarne D.; Orecchia P.; Palomba D.; Pasa S.; Patrone E.; Ponassi R;Scapolla A.; Tortolina L.; Vasile F.; Mazzei M.; Parodi S. (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#url
  • http://www.fasebj.org/content/early/2005/03/25/fj.04-2369fje (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroVolume
  • 19 (literal)
Rivista
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  • 26 (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroFascicolo
  • 1 (literal)
Note
  • ISI Web of Science (WOS) (literal)
  • Scopu (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#affiliazioni
  • ISMAC-CNR Genova: Cristina D'Arrigo; ELIGIO PATRONE; UNI. GE, Medicina, IST GE, Inst Giannina Gaslini GE, Xeptagen SpA, Naples: Altri (literal)
Titolo
  • Sequence specific peptidomimetic molecules inhibitors of a protein-protein interaction at the helix 1 level of c-myc (literal)
Abstract
  • Our work is focused in the broad area of strategies and efforts to inhibit protein - protein interactions. The possible strategies in this field are definitely much more varied than in the case of ATP-pocket inhibitors. In our previous work ( 10), we reported that a retro-inverso ( RI) form of Helix1 (H1) of c-Myc, linked to an RI-internalization sequence arising from the third alpha-helix of Antennapedia (Int) was endowed with an antiproliferative and proapoptotic activity toward the cancer cell lines MCF-7 and HCT-116. The activity apparently was dependent upon the presence of the Myc motif. In this work, by ala-scan mapping of the H1 portion of our molecules with Daa, we found two amino acids necessary for antiproliferative activity: D-Lys in 4 and D-Arg in 5 ( numbers refer to L-forms). In the natural hetero-dimer, these two side chains project to the outside of the four alpha-helix bundle. Moreover, we were able to obtain three peptides more active than the original lead. They strongly reduced cell proliferation and survival (RI-Int-VV-H1-E2A, S6A, F8A; RI-Int-VV-H1-S6A, F8A, R11A; RI-Int-VV-H1-S6A, F8A, Q13A): after 8 days at 10 muM total cell number was similar to 1% of the number of cells initially seeded. In these more potent molecules, the ablated side chains project to the inside in the corresponding natural four alpha-helix bundle. In the present work, we also investigated the behavior of our molecules at the biochemical level. Using both a circular dichroism (CD) and a fluorescence anisotropy approach, we noted that side chains projecting at the interior of the four alpha-helix bundle are needed for inducing the partial unfolding of Myc-H2, without an opening of the leucine zipper. Side chains projecting at the outside are not required for this biochemical effect. However, antiproliferative activity had the opposite requirements: side chains projecting at the outside of the bundle were essential, and, on the contrary, ablation of one side chain at a time projecting at the inside increased rather than decreased biological activity. We conclude that our active molecules probably interfere at the level of a protein - protein interaction between Myc-Max and a third protein of the transcription complex. Finally, CD and nuclear magnetic resonance (NMR) data, plus dynamic simulations, suggest a prevalent random coil conformation of the H1 portion of our molecules, at least in diluted solutions. The introduction of a kink ( substitution with proline in positions 5 or 7) led to an important reduction of biological activity. We have also synthesized a longer peptido-mimetic molecule (RI-Int-H1-S6A, F8A-loop-H2) with the intent of obtaining a wider zone of interaction and a stronger interference at the level of the higher-order structure (enhanceosome). RI-Int-H1-S6A, F8A-loop-H2 was less active rather than more active in respect to RI-Int-VV-H1-S6A, F8A, apparently because it has a clear bent to form a - sheet ( CD and NMR data). (literal)
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