MOLECULAR

CHAPERONES

Biochemistry
1st year Medicine - Section E2
Group members:
David, Brian
Dela Cruz, Jonella Ruth
Dinzon, Aira Mae
Dizon, Fideline
Esguerra, Celina

Proteins

Proteins are any large group of nitrogenous compounds of high molecular weight that are essential constituents of
all living organisms. They consist of one or more chains of amino acids linked by peptide bonds and are folded
into a specific three-dimensional shape maintained by further chemical bonding.

Funtions of proteins
 Transport proteins
 Hormones
 Catalyst / Enzymes
 Receptor / Cell signals

Examples of proteins

Fibrous Proteins- from muscle fiber, tendons, connective tissues and bones.

-e.g. Actin, Collagen

Globular Proteins- are more water soluble than the other classes of proteins and they have several functions
including transporting, catalyzing and regulating.

-e.g Albumin, Hemoglobin

Membrane Proteins-relaying signals within cells, allowing cells to interact, and transporting molecules
 -Glucose transporter, Potassium Channel

Protein synthesis

a process of creating protein molecules.

Two process to manufacture proteins

1. Transcription- First synthesizes as RNA molecule that is complementary to one strand of the DNA double
helix for a particular gene. The RNA copy is taken out of the nucleus and into the cytoplasm

2. Translation- the information in the RNA is used to manufacture protein by aligning and joining specific
amino acids, then the protein folds into a specific three dimensional form necessary for its function.

Protein Folding

Physical process by which a polypeptide folds into characteristics and functional three-dimensional structure from
unfolded polypeptide after being translated from a sequence of mRNA to a linear chain of amino acids.

Protein Misfolding

− Failure of a protein to achieve its native conformation misfolded or incompletely folded proteins.

− Failure to maintain that conformation due to instability as a result of environmental change or mutation.

− Common at elevated temperatures and under stressful conditions

- Protein misfolding is the molecular basis of a wide range of human diseases, including the amyloidoses.
Commonly results to neurologic diseases

Protein Aggregates

Moreover, many proteins fail to spontaneously refold in vitro. Instead they form insoluble aggregates,
disordered complexes of unfolded or partially folded polypeptides held together predominantly by hydrophobic
interactions. Aggregates represent unproductive dead ends in the folding process. Cells employ auxiliary proteins
to speed the process of folding and to guide it toward a productive conclusion.
Proteins are conformationally dynamic molecules that can fold and unfold hundreds or thousands of times in their
lifetime. How do proteins, once unfolded, refold and restore their functional conformation? First, unfolding rarely
leads to the complete randomization of the polypeptide chain inside the cell. Unfolded proteins generally retain a
number of contacts and regions of the secondary structure that facilitate the refolding process. Second,
chaperone proteins can “rescue” unfolded proteins that have become thermodynamically trapped in a misfolded
dead end by unfolding hydrophobic regions and providing a second chance to fold productively.

Molecular chaperones stabilize unfolded or partially folded proteins. Chaperones are required for the correct
targeting of proteins to their subcellular locations.

Protein denaturation
A loss of three dimensional structure sufficient to cause loss of funtion
 Heat
 Extremes of pH

Refolding of proteins

 Denatured proteins can renature spontaneously to form biologically active protein.

 Chaperones capture unfolded polypeptides, stabilize intermediates and prevent misfolded species from
accumulating in stressed cells.
Molecular chaperones

A large group of unrelated protein families that are also known as heat shock proteins or stress
proteins. Whose role is to interact and stabilize partially folded or improperly folded polypeptides, It
also facilitates correct folding pathways or provide microenvironment in which folding can occur.

Any protein that interacts with, stabilizes or helps another protein to acquire it’s functionally active
confirmation, without being present in it’s final structure.

Role of molecular chaperones

 Prevents inappropriate association or aggregation.

 Facilitates correct protein folding.
 Act as a catalyst.
 Rescue proteins.

Properties of molecular chaperones

 Some chaperones are non-specific, and interact with wide variety of polypeptide chains, but
others are restricted to specific target.
 Binds predominantly to hydrophobic regions of unfolded proteins and prevent their aggregation.
 They often couple ATP binding/ hydrolysis to the folding process.
 Inducible by conditions that cause unfolding of newly synthesized proteins.
 Interaction found in various cellular compartments.
 Do not interact with native proteins, nor do they form part of the final structure.

Molecular Chaperones ( Heat Shock Proteins)

HSP 60
 Also known as Chaperonins
 Consist of 2 distinct subgroups
- Group I chaperonins (GroEL/GroES) found in prokaryotes and endosymbiotic organelles such as Mitochondria
- Group II chaperonins (TRiC/ CCT) present in archaea and cytosolic compartment of eukaryotic systems
 Transportation and refolding of proteins from cytoplasm to mitochondrial matrix
 Assist in folding linear amino acid chains to respective three dimensional structure
 Structure- Hollow cylinder consisting of 2 rings with 7 subunits stacked back to back with hydrophobic
residues in either end of the cylinder
HSP 70
 Translocation across organelle membranes - assist in protein transport into mitochondria and endoplasmic
reticulum
 Disaggregation of aggregates
 Protects protein under stress
 Composed of two major functional domains:
- N - Terminal (ATPase domain)
- C - Terminal ( Polypeptide bonding)
 Stabilizes proteins prior to complete folding

Other molecular chaperones

HSP 25
 Binds and stabilize denatured protein under conditions of cellular stress, ageing and degenerative diseases
 It has 24 subunits, each with immunoglobulin fold arrange as hollow shell with holes ( resembles a soccer
ball)

HSP 90
 Binding and stabilization - stabilizes proteins prior to complete folding or activation
 Regulation of steroid receptors, protein kinase
 Forms a homodimer through its C-terminal residues
 Forms stable complexes with inactive glucocorticoid receptor and other transcript factors

HSP 100
 Hexameric structure
 Thermotolerance
 Resolubilization of aggregates
 Facilitates proteolysis

Summary of Molecular chaperones

Molecular chaperones Function Structure
HSP 60 Protein folding Hollow cylinder consisting of 2 rings
with 7 subunits stacked back to
back with hydrophobic residues in
either end of the cylinder
HSP70 Membrane transport of two major functional domains, N -
proteins, protein unfolding Terminal (ATPase domain) C -
Terminal ( Polypeptide bonding)
HSP 25 Stabilization of misfolded 24 subunits, each with
proteins immunoglobulin fold arrange as
hollow shell with holes ( resembles
a soccer ball)
HSP 90 Binding and stabilization, Forms a homodimer through its C-
regulatory interactions with terminal residues
signaling proteins
HSP 100 Protein disaggregation, Hexameric structure
thermotolerance

Diseases resulting from defective molecular chaperones

Amyloid diseases
 Formation of long, fibrillar protein consisting of Beta pleated sheets.
 Parkinsons
 Hungtintons
 Alzheimer disease

Prion Diseases

 Prion protein is the causative agent of transmissible spongiform encephalopathies

 Creutzfeldt- jakob disease-humans
 Scarpie- sheeps
 Mad cow disease- cattles

Drugs for specific chaperone activities

 Drugs targeting specific chaperone activities Immunophilins HSP70 HSP90

 15- Deoxyspergualin (DSG) binds to HSP70 class
 Immunosuppresive and cytotastic
 Allows reverse renal allograft rejection
 Useful in combunation-theraphy after solid- organ transplant
 Non steroidal anti inflammatory drugs (NSAIDS)
 Enhance chaperone activity
 Mechanism for inducing chemical chaperones allows for efficient correction/ prevention if misfolds
 Potentiate HSP 1 activation

Induction of protein and chemical chaperones

 Induction of proteins and chemical chaperones has been long recognized that heat treated or chemically
stressed cells and tissues gain a tolerance to further stressful insult.
 This condition results in large part from the induction of hsps and chemical chaperones that enhance the
cellular environment for protein folding and stability. A beat shock response is observed in many tissues
after an elevation of body temperature and fever may in part be an adaptation to naturally increase
intracellular chaperone activity
As an immunologic adjuvant

 A basic intracellular function of Hsps is peptide chaperoning within and across cellular compartments.
Hsp70 members may chaperone potential MHC class I
 Reduce immunological variance of disease with HSP peptide complex in vivo
 Against tumors: may obtain entire antigenic receptor tumor , effective with specific T cell response to
disease cells.