Circular Dichroism sessions


CD1: CD Principles and instrumentation

  • CD lecture handouts here (password: CD2017)
  • How to study proteins by circular dichroism - Kelly BBA 05 (Paper) 
  • Textbook 'Circular Dichroism and the Conformational Analysis of Biomolecules' (for further reading)

CD2: Protein analysis by CD – secondary structure determination and protein stability

  • In this experimental session we will record and analyse far-UV CD spectra of proteins/peptides (Myoglobin and poly-Lysine). Stock protein solutions will be available
  • Using the Jasco J-815 Circular Dichroism Spectropolarimeter > general setup protocol here
  • Ref: Using circular dichroism spectra to estimate protein secondary structure (Nature Protocols here)
  • Ref: Circular Dichroism (CD) Spectroscopy - Practical Protein CD (here)
  • Homework: Students/group should analyse data and prepare a figure with CD spectra of poly-lysine at basic and neutral conditions and use the Myoglobin spectra to estimate secondary structure, as per references above (e.g. using Bestsel)

CD3-MS3: Research case study: CD analysis of Parkinson’s Disease alpha-synuclein

  • α‑synuclein (α‑syn) plays important roles in health and disease (Parkinson's Disease and Lewi body dementias) and its nature as an intrinsically disordered protein has been difficult to reconcile with its unfolded structure; recently it was proposed that native α‑syn exists as a folded tetramer versus an unfolded peptide and there is ongoig debate on this matter.  Here we will use freshly prepared recombinant human alpha-synuclein for far-UV CD and MS analysis to investigate its structure and folding. Students must get acquainted with the discussion and prepare this lab course by reading the papers in this ZIP file.
  • Due to availability of freshly prepared α‑syn, this experimental session will take place for all students only on  the 17th November; at 14:00 half of the students start at the CD lab and the other half starts at the MS lab and then the groups switch during the afternoon.

Anexos