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Lecture - 18-Protein Folding, Chaperones, Prions, Alzyheimer's
Lecture - 18-Protein Folding, Chaperones, Prions, Alzyheimer's
Protein Folding
Molecular Chaperones
Prions
Alzyheimer’s
Tertiary Structure of Proteins
Two methods: 1. X-RAY diffraction crystal structure
2. NMR solution structure
From the diffraction pattern (spots and intensity) one can get a
mathematical description of the electron density of a molecule. With
proper model construction a 3-D image of the protein is constructed.
NMR
By using chemical shifts of backbone hydrogens and their
chemical splitting bond angles can be determined. COSY NMR
or Correlated Spectroscopy. By manipulating parameters
protons that are close to each other in space but not linked
through bonds can be determined by NOSY NMR or Nuclear
Overhauser spectroscopy. Growing the protein in bacteria where
the carbon source can be substituted by 13C and the nitrogen by
15
N (stable isotope substitution) more restraints can be achieved.
Alan Fersht
protein folding can be seen as a connection
between the genome (sequence) and what the
proteins actually do (their function).
Protein folding problem
• For
ΔGD-N = ΔHD-N - TΔSD-N
• Gives
ΔGD-N(T2) = ΔHD-N(T1) + ΔCp(T2 – T1)- T2(ΔSD-N(T1) + ΔCpT2 / T1)
D <—> N
Folding of a 20-mer poly Ala
Unfolding of the DNA Binding Domain of
HIV Integrase
Two state transitions in multi-state reactions
Rate determining steps
Energy profiles during Protein Folding
Theories of protein folding
• N-terminal folding
• Hydrophobic collapse
• The framework model
• Directed folding
• Proline cis-trans isomerisation
• Nucleation condensation
Molecular Chaperones
D<---->N
Ag
Molecular Chaperone Function
• Disulfide isomerases
• Peptidyl-prolyl isomerases (cyclophilin, FK506)
• Bind the denatured state formed on ribozome
• Heat shock proteins Hsp (DnaK)
• Protein export & delivery SecB
What happens if proteins don't fold correctly?
• Positive cooperativity
• Multiple binding sites
Allosteric nature of GroEL
GroEL changes affinity for denatured proteins