A chemical approach to the structural and biophysical study of the Id proteins and to the modulation of their biological functions

• Applicant: Professorin Dr. Chiara Cabrele
• Subject Area: Biological and Biomimetic Chemistry
• Term: from 2003 to 2010
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 5396788

• Final Report Year: 2011

Final Report Abstract

The Id proteins (Id1-4) belong to the large family of helix-loop-helix (HLH) transcription factors and act as inhibitors of DNA binding and cell differentiation. They are important during development, cell fate and lineage determination, neuro- and angiogenesis. Also, upregulated Id protein expression has been observed in several cancer types as well as in vascular disorders, and has been related to tumor angiogenesis, migration and invasion. Therefore, the Id proteins represent potential therapeutic targets. We have exploited a chemical approach for the synthesis, structural characterization and function control of the Id protein family. Large fragments of the four Id proteins, including amino acid substituted and fluorescence-labeled analogs, as well as the full-length Id3 protein were prepared by solid-phase technique and investigated by spectroscopic methods. Moreover, a peptide consisting of a dimeric form of a short sequence of the Id1 HLH motif has displayed inhibitory activity on proliferation and migration of vascular smooth muscle cells, while inducing differentiation. This may have an impact on pathological phenotypes such as atherosclerosis and hypertension. Our synthetic, conformational and Id-protein modulation studies are still ongoing and aim at the accomplishment of those tasks that could not be reached during the granted period. In particular, future work will focus on the total chemical synthesis of Id1 and Id2, which, beside Id3, are the biologically and pathologically most relevant members of the Id family, on the understanding of their folding and association properties and on the development of Id-protein inhibitors for the modulation of cell differentiation, proliferation, and migration in pathological systems.

Publications

• Synthesis and Conformational Properties of Protein Fragments Based on the Id Family of DNA-Binding and Cell-Differentiation inhibitors. Biopolymers - Peptide Science, Vol. 80. 2005, Issue 6, pp. 762–774.
  S. Kiewitz, C. Cabrele
  
• A Short Id2 Protein Fragment Containing the Nuclear Export Signal Forms Amyloid-Like Fibrils. Biochemical and Biophysical Research Communications, Vol. 346. 2006, Issue 1, pp. 182–187.
  N. Colombo, J. Schroeder, C. Cabrele
  
• An Improved Synthesis of 3,4-(Aminomethano)proline and its Incorporation into Small Oligopeptides. European Journal of Organic Chemistry, Vol. 2006, Issue 19, pp. 4440–4450.
  F. Brackmann, N. Colombo, C. Cabrele, A. de Meijere
  
• Chemoenzymatic resolution of epimeric cis 3-carboxycyclopentylglycine derivatives. Tetrahedron, Vol. 62. 2006, Issue 15, pp. 3502–3508.
  C. Cabrele, F. Clerici, R. Gandolfi, M. L. Gelmi, F. Molinari, S. Pellegrino
  
• Fluorescence- and Luminescence-based Methods for the Determination of Affinity and Activity of Neuropeptide Y Y2 Receptor Ligands. European Journal of Pharmacology, Vol. 551. 2006, Issue 1–3, pp. 10–18.
  R. Ziemek, A. Brennauer, E. Schneider, C. Cabrele, A. G. Beck-Sickinger, G. Bernhardt, A. Buschauer
  
• Stepwise Solid-Phase-Synthesis and Spontaneous Homodimerization of the Helix-Loop-Helix Protein Id3. ChemBioChem, Vol. 7. 2006, Issue 8, pp. 1164–1168.
  J. Svobodová, C. Cabrele
  
• Synthesis and Conformational Analysis of Id2 Protein Fragments: Impact of Chain Length and Point Mutations on the Structural HLH Motif. Journal of Peptide Science, Vol. 12. 2006, Issue 8, pp. 550–558.
  N. Colombo, C. Cabrele
  
• Determination of affinity and activity of ligands at the human neuropeptide Y Y4 receptor by flow cytometry and aequorin luminescence. Journal of Receptors and Signal Transduction, Vol. 27. 2007, Issue 4, pp. 217-233.
  R. Ziemek, E. Schneider, A. Kraus, C. Cabrele, A. G. Beck-Sickinger, G. Bernhardt, A. Buschauer
  
• γ-Aminoadamantanecarboxylic Acids Through Direct C–H Bond Amidations. European Journal of Organic Chemistry, Vol. 2007, Issue 9, pp. 1474–1490.
  L. Wanka,C. Cabrele, M. Vanejews, P. R. Schreiner
  
• Recognition of the Helix-Loop-Helix Domain of the ld Proteins by an Artificial Luminescent Metal Complex Receptor. Journal of Molecular Recognition, Vol. 21. 2008, Issue 2, pp. 79–88.
  S. D. Kiewitz, M. Kruppa, A. Riechers, B. König, C. Cabrele
  
• Switching from the Unfolded to the Folded State of the Helix-Loop-Helix Domain of the Id Proteins Based on the O-Acyl Isopeptide Method. Journal of Peptide Science, Vol. 14. 2008, Issue 11, pp. 1209–1215.
  S. D. Kiewitz, T. Kakizawa, Y. Kiso, C. Cabrele
  
• Conformation and Stability of the Helix-Loop-Helix Domain of the Id Protein Family. In: M. Lebl Ed., Peptides: Breaking Away: Proceedings of the 21st American Peptide Symposium, American Peptide Society, Bloomington, Indiana, ISBN 0-9715560-3-2, 2009, pp. 343-344.
  S.D. Kiewitz, Y. Kiso, C. Cabrele
  
• Stable Right- and Left-handed Peptide Helices Containing Cα-tetrasubstituted α-Amino Acids. Journal of Organic Chemistry, Vol. 74. 2009, Issue 10, pp. 3718–3726.
  A. A. Grauer, C. Cabrele, M. Zabel, B. König
  
• Synthetic Peptides Containing a Conserved Sequence Motif of the Id Protein Family Modulate Vascular Smooth Muscle Cell Phenotype, Bioorganic & Medicinal Chemistry Letters, Vol. 19. 2009, Issue 22, pp. 6298–6302.
  S. Pellegrino, N. Ferri, N. Colombo, E. Cremona, A. Corsini, R. Fanelli, M. L. Gelmi, C. Cabrele
  
• G protein-coupled receptors function as logic gates for nanoparticle binding and cell uptake. Proceedings of the National Academy of Sciences of the USA, vol. 107 no. 23, pp. 10667–10672.
  W. Hild, K. Pollinger, A. Caporale, C. Cabrele, M. Keller, N. Pluym, A. Buschauer, R. Rachel, J. Tessmar, M. Breunig, A. Goepferich
  
• Peptide-Based Modulation of Helix-Loop-Helix Proteins. Abstracts of Papers, 239th ACS National Meeting, San Francisco, CA, USA, ORGN-478, ISBN 9780841225732.
  C. Cabrele, S. Pellegrino, N. Ferri, M. L. Gelmi, M. Beißwenger
  
• Side-Chain Cyclization Based on Serine Residues: Synthesis, Structure and Activity of a Novel Cyclic Analogue of the Parathyroid Hormone Fragment 1-11. Journal of Medicinal Chemistry, Vol. 53. 2010, Issue 22, pp. 8072–8079.
  A. Caporale, M. Sturlese, L. Gesiot, F. Zanta, A. Wittelsberger, C. Cabrele
  
• Synthesis and Conformation of an Analog of the Helix-Loop-Helix Domain of the Id1 Protein Containing the O-Acyl Iso-Prolyl-Seryl Switch Motif. Journal of Peptide Science, Vol. 16. 2010, Issue 6, pp. 303–308.
  M. Beißwenger, T. Yoshiya, Y. Kiso, C. Cabrele
  
• Functional Reconstitution of Human Neuropeptide Y (NPY) Y2 and Y4 Receptors in Sf9 Insect Cells. Journal of Receptors and Signal Tranduction, Vol. 31. 2011, No. 4 , pp. 271-285.
  N. Pop, P. Igel, A. Brennauer, C. Cabrele, G. Bernhardt, R. Seifert, A. Buschauer
  
• Wenn Proteine ihren Partner wechseln. BIOspektrum, Vol. 17. 2011, Issue 1, pp 12-16.
  C. Cabrele