MOLECULAR

CHAPERONES

Group 5 – A2
Capacio, Diane Chloe
Cardeño, Sheree Ann Rachille
Casem, Princess Joy
Casteñeda, Felix II
Castro, John Michael
Ceneta, Raymond
 INTRODUCTION
Chaperones
 Proteins which assist in protein folding
 Assists in the translocation of polypeptide
chains
 Multiprotein assembly and disassembly
 Escort some proteins to specific
destinations
 Rescue proteins trapped in misfolded
dead-ends (folded in their hydrophobic
regions)
 Protein folding
 During the synthesis of proteins, parts of it
already begin folding
 The instruction on the sequence’s proper
folding is dependent on the amino acid
sequence itself
Protein folding
 Most of the hydrophobic amino acids are
buried inside the structure (core)
 Some proteins already fold as it comes
out of the ribosome in a matter of
seconds, forming what is called a molten
globule
Chaperones assist in folding of some
proteins
Chaperones assist in folding of some
proteins
 They are of much help since there are a
number of pathways at which the protein
can achieve its final form from an
unfolded state
 Many of these chaperones are termed
heat shock proteins because they are
made in abundance at an elevated
temperature (~42oC for cells normally at
37oC)
Chaperones assist in folding of
some proteins
 A feedback-like system is made in this
setting, since an increase in unfolded
proteins resulting from an increased
temperature will eventually lead to the
synthesis of chaperones
Chaperones assist in folding of
some proteins
 Some of the major chaperone families
include Hsp60 and Hsp70, which
functions at different organelles
 The mitochondria have its own set of
Hsp60 and Hsp70 proteins which are
distinct from those found in the cytosol
and BIPs (special types of Hsp70) in the
endoplasmic reticulum
Chaperones assist in folding of
some proteins
 Hsp70s act early in the synthesis of the
protein before that protein leave the
ribosome
 Hsp60s, however, acts on the fully
synthesized protein by forming a barrel-
like structure. Hsp60s are also termed
as “Chaperonin” and they have what is
called an “isolation chamber” where the
misfolded protein is fed into
Chaperones assist in folding of
some proteins
 Chaperones use a series of ATP
hydrolysis in aiding a protein to fold
correctly
How does the cell recognize
misfolded proteins?
 Usually, when a hydrophobic region is at
the protein globule surface, it is
considered abnormal
 Presence of these kinds of proteins may
render, not only nothing for the cell, but
also danger. Aggregations of misfolded
proteins may result to some known
severe human diseases
Types of Molecular Chaperones
 Folding
 Holding
 Those that increase solubility of proteins
Role in protein assembly
 They guide in the proper folding of
proteins
 Misfolded proteins are allowed to refold
into their correct conformation by having
a suitable environment which is provided
by the chaperones, chaperonins in
particular
Refolding of denatured proteins
 In a study involving recombinant HSP17
of Synechocystis sp., malate
dehydrogenase which underwent
denaturation made complexes with
HSP17 which then functioned as a
storage of the unfolded protein, and later
on transported to the network of
Dnak/DnaJ/GrpE and GroEL/ES
chaperones for eventual refolding (Török
etal., 2001)
Production of recombinant proteins
 In a study using E. coli, overproduction
of bacterial chaperones, GroEL and
GroES, via genetic engineering resulted
to high yield of recombinant human
protein-tyrosine kinase p50csk which
are in an insoluble forms within the
bacterial cell.
Production of recombinant proteins
 Overproduction of these chaperones
resulted in the subsequent dissolution of
the protein kinase and enabled its
purification (Amrein et al., 1995)
Production of cancer vaccines
 HSPs which are derived from a
particular tumor has been known to
bring on immunity against the tumor
where came from that tumor. Vaccines
of this kind are effective on different
types of tumors and can bypass the
need for identifying the responsible
peptide which causes the immunity
Production of cancer vaccines
 These vaccines are specific for the
tumor and for the patient because they
confine the identity of the tumor cell.
HSP96 has already been studied with a
number of cancer illnesses such as,
melanoma, renal cell carcinoma, gastric
cancer, pancreatic cancer, low-grade
lymphoma, colorectal cancer and
chronic myelogenous leukemia (Oki and
Younes, 2004)
Possible causes of Protein
Misfolding

•Genetic Mutation
•Increased Heat Production
Properties of Chaperones
•Present in a wide range of
species from bacteria to
humans
•Many are so called Heat
Shock Proteins
•Some are inducible by
conditions that cause unfolding
of newly synthesized proteins
•They bind to predominantly
hydrophobic regions of
unfolded proteins and prevent
their aggregation
•They act in part as quality
control
•Associated with ATPase
activity
•Found in cytosol, mitochondria
and lumen of ER
Activation of Chaperones
 Rely on ATP-driven conformational
change

Most of the hydrophobic amino acids

are buried inside the structure (core)
 Holding Chaperones
 Maintains partially folded protein
on their surface to await
availability of folding chaperone
upon stress abatement
Chaperons w/c Promotes Solubility
of Protein

 Aggregates as a result of stress

2 Major Types of Molecular
Chaperones
 Hsp 6O
Also called Chaperonin
Barrel like structure- where unfolded
protein is retained, giving it time &
suitable env’t to fold properly
Hsp 7O

 These protein acts early, recognizing

small strech of hydrophobic amino acid
pn a protein surface
GroEL-GroES System
 It is a bacterial chaperone belong to Hsp
6O family
 It is a hollow cylinder consist of 2 seven
subunit rings stacked back to back
Dnak Chaperones
 They are chaperones of host protein,
refolding, translocation and general
mngt of deleterious effest of stress
 They are under the hsp70 family
Hsp33
 Also know as REDOX-ACTIVATED
HOLDING CHAPERONES
 It is a redox regulated chaperon that
stabilized protein unofolded by severe
oxidative stress
Hsp31
 Also known as Managing Heat &
AcidStress
 It fxn as a holding chaperons that helps
cell handle protein unfolding under
extremes of temp.
 Plays role in ability of strved E. Coli to
survive in a acid stress
Clp Disaggregase
 Belong to hsp100 family w/c role is to
degrade unfolded/misfolded protein or to
stabilized and reactivate aggregate
proteins
PROSPECTIVE THERAPEUTIC
RATIONALES THAT INVOLVE
CHAPERONES
 Drugs targeting specific
Chaperone Activities

1. Immunophilins
a. Immunosuppressants cyclosporin A
and FK506
• Transplantation procedure,
autoimmune and inflammatory
diseases.
• Useful as neurotrophic agents.
2. HSP70
a. DSG interact selectively with HSP70
class chaperones
• Potent immunosuppressive agent.
• Maybe able to reverse renal
allograft rejection episodes.
• Maybe quite useful in combination-
therapy.
• Further studies required.
3. HSP90
• Inhibitors of signal transduction
• Examples are GA and
benzoquinone ansamycins which
are potential as anti cancer
agents
Induction of protein and chemical
chaperones
 Injury protection
•Induction of heat shock response
might be an effective prophylactic
treatment to minimize myocardial
injury.

•Herbimycin which induce activation of

HSF1 has been shown to protect
cardiomyocytes in culture.
References
Campbell, NA, JB Reece, LA Urry, ML Cain, SA Wasserman, PV Minorsky and RB
Jackson. 2008. Biology 8th ed. Pearson Benjamin Cummings, CA
Alberts, B, A johnson, J Lewis, M Raff, K Roberts and P Walter. 2008. Molecular Biology of
the Cell 5th ed. Garland Science, UK
Amrein, KE, B Takacs, M Steiger, J Molnos and P Burn. 1995. Purification and
characterization of recombinant human p50csk protein-tyrosine from an Escherichia
coli expression system overproducing the bacterial chapreones GroES and GroEL.
Proc. Nat. Acad. Sci. 92(4):1048-1052.
Török, Z, P Goloubinoff, I Horvath, NM Tsvetkova, A Glatz, G Balogh, V Varvasovszki, DA
los, E Veirling, JH Crowe and L Vigh. 2001. Synechocystis HSP17 is an amphitropic
protein that stabilize heat-stressed membranes and binds denatured proteins for
subsequent chaperone-mediated refolding. Proc. Nat. Acad. Sci. 98(6):3098-3103.
Oki, Y and A Younes. 2004. Heat shock protein-based cancer vaccines.
Expert Review of Vaccines 3(4):403-411.
http://faculty.washington.edu/baneyx/Chaperones/Chaperones.html
Smith, D, L Whitesell and E Katsanis. 1999. Molecular Chaperones: Biology and Prospects
for Pharmacological Intervention. Pharmacological Reviews 50(4): 493-514.