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Protein Aggregation
Protein Aggregation
Stress (environmental)
induced misfolding generates
sticky aggregation prone
conformation
Normally
folded
protein
Oligomers
Diseases
Protein involved
Alzheimers disease
Amyloid-
Parkinson disease
-Synuclein
Diabetes type 2
Amylin
Superoxide dismutase
Hemoglobin
Hungtington disease
Huntingtin
Creutzfeldt-Jakob disease
Prion protein
Amyloidosis
2. Loss of function
Other effect of the misfolded protein may be due to loss of function, as observed
in cystic fibrosis. There is a mutation in the CFTR sequence
3. Accumulation
Protein aggregates are sometimes converted to a fibrillar structure. Fibrils
themselves are not toxic but insoluble. Their accumulation cause tissue damage
(amyloidoses)
Polyethylene glycol
(PEG 3350)
0.1-0.4 g/L
L-Arginine hydrochloride
0.4-0.8M
Nondenaturating
concentrations of Urea
< 2.0 M
K-Glutamate
~5M
Nondenaturating
< 1.0 M
concentrations Gdm/ClH
Proline
~1M
Methylurea
1.5-2.5 M
Glycerol
20-40 %
Ethylurea
1.0-2.0 M
Sorbitol
20-30 %
Formamide
2.5-4.0 M
Sucrose
~1M
Methylfomamide
2.0-4.0 M
Trehalose
~1M
Acetamide
1.5-2.5 M
TMAO
(trimethylamine N-oxide)
~1M
Ethanol
Up to 25%
Sulfo Betaine
~1M
(http://www.ls.huji.ac.il/~purification)
n-Penthanol
1.0-10.0 mM
Lauryl Maltoside
0.06 mg/ml
n-Hexanol
0.1-10.0 mM
CETAB
0.6 mg/ml
Cyclohexanol
0.01-10.0 mM CHAPS
10-60 mM
Tris
> 0.4 M
Triton X-100
10 mM
Dodecyl Maltoside
2.0-5.0 mM
Cyclodextrin
Sarkosyl
0.05-0.5 %
20-100 mM
(http://www.ls.huji.ac.il/~purification)
Chaperones
Thermophilic Proteins
Living organisms can be found in the most unexpected places,
including deep sea vents at > 100 C and several hundred bars
pressure, in hot springs, and most recently, deep in the bowels of
the earth, living off H2 formed by chemical decomposition of rocks!
The proteins found in thermophilic species are much more stable
than their mesophilic counterparts (although this corresponds to
only 3 - 8 kcal/mol of free energy).
The upper limit of temperature growth for bacteria is about 110
C.
Many of the species found in these extreme environments
(T > 100C, pH 2) belong to the Archeae kingdom.
Protein Stability
Protein stability is the net balance of forces,
which determine whether a protein will be in
its native folded conformation or a denatured
state.
Chemical Stability
Chemical stability involves loss of integrity due to bond
cleavage.
Protein Stability
Protein stability is important for many reasons:
Providing an understanding of the basic thermodynamics of
the process of folding,
increased protein stability may be a multi-billion dollar value
the in food and drug processing, and in biotechnology and
protein drugs.