Heat Sock Proteins

Heat shock proteins (HSP) are a group of proteins induced by heat shock or the expression of
which is increased when the cells are exposed to elevated temperatures (or stress conditions),
the most prominent members of this group are a class of functionally related proteins involved
in the folding and unfolding of other proteins.
Heat-shock proteins (HSPs), or stress proteins, are highly conserved and present in all
organisms and in all cells of all organisms. Selected HSPs, also known as chaperones, play
crucial roles in folding/unfolding of proteins, assembly of multi-protein complexes,
transport/sorting of proteins into correct subcellular compartments, cell-cycle control and
signaling, and protection of cells against stress/apoptosis.
Heat shock proteins are present in cells under normal conditions, but are expressed at high
levels when exposed to a sudden temperature jump or other stress. Heat shock proteins
stabilize proteins and are involved in the folding of denatured proteins. High temperatures and
other stresses, such as altered pH and oxygen deprivation, make it more difficult for proteins to
form their proper structures and cause some already structured proteins to unfold. Left
uncorrected, mis-folded proteins form aggregates that may eventually kill the cell. Heat Shock
Proteins are induced rapidly at high levels to deal with this problem. Increased expression of
HSPs is mediated at multiple levels: mRNA synthesis, mRNA stability, and translation efficiency.
Most heat shock proteins are molecular chaperones. Chaperones aid in the transport of
proteins throughout the cell's various compartments.

The enhanced synthesis of a few proteins immediately after subjecting cells to a stress such as
heat shock was first reported for drosophila cells in 1974, and the universality of the response
from bacteria to human was recognized shortly thereafter. Recently, the emphasis in this field
has focused on the unction of various heat shock proteins and their possible role as molecular
chaperones.

Functions of Heat Shock Proteins:

Upregulation in stress
Production of high levels of heat shock proteins can also be triggered by exposure to different
kinds of environmental stress conditions, such as infection, inflammation, exercise, exposure of
the cell to toxins (ethanol, trace metals, and ultraviolet light etc.), starvation, hypoxia, nitrogen
deficiency (in plants), or water deprivation. As a consequence, the heat shock proteins are also
referred to as stress proteins and their upregulation is sometimes described more generally as
part of the stress response.
The mechanism by which heat-shock (or other environmental stressors) activates the heat
shock factor has been determined in bacteria. During heat stress, outer membrane proteins
(OMPs) do not fold and cannot insert correctly into the outer membrane. They accumulate in
the periplasmic space. These OMP's are detected by DegS, an inner membrane protease, that
passes the signal through the membrane to the sigmaE transcription factor. However, some
studies suggest that an increase in damaged or abnormal proteins brings HSPs into action.
Role as chaperone
Several heat shock proteins function as intra-cellular chaperones for other proteins. They play
an important role in protein-protein interactions such as folding and assisting in the
establishment of proper protein conformation and prevention of unwanted protein
aggregation. By helping to stabilize partially unfolded proteins, HSPs aid in transporting proteins
across membranes within the cell.
Some members of the HSP family are expressed at low to moderate levels in all organisms
because of their essential role in protein maintenance.
Management
Heat-shock proteins also occur under non-stressful conditions, simply "monitoring" the cell's
proteins. Some examples of their role as "monitors" are that they carry old proteins to the cell's
"recycling bin" (proteasome) and they help newly synthesized proteins fold properly.

These activities are part of a cell's own repair system, called the "cellular stress response" or
the "heat-shock response".
Cardiovascular
Heat shock proteins appear to serve a significant cardiovascular role. Hsp90, hsp84,
hsp70, hsp27, hsp20, and alpha B crystallin all have been reported as having roles in the
cardiovasculature.
Hsp90 binds both endothelial nitric oxide synthase and soluble guanylate cyclase, which in turn
are involved in vascular relaxation.
A kinase of the nitric oxide cell signaling pathway, protein kinase G, phosphorylates a small heat
shock protein, hsp20. Hsp20 phosphorylation correlates well with smooth muscle relaxation
and is one significant phosphoprotein involved in the process. Hsp20 appears significant in
development of the smooth muscle phenotype during development. Hsp20 also serves a
significant role in preventing platelet aggregation, cardiac myocyte function and prevention of
apoptosis after ischemic injury, and skeletal muscle function and muscle insulin response.
Hsp27 is a major phosphoprotein during a woman's contractions. Hsp27 functions in small
muscle migrations and appears to serve an integral role.
Immunity
Extracellular and membrane bound heat-shock proteins, especially Hsp70 are involved in
binding antigens and presenting them to the immune system.

The initial studies on cloned heat shock genes and purified proteins led to two important
results. First, the heat shock proteins proved to be incredibly highly conserved among widely
divergent organisms. For example, the major heat shock protein, hsp70, has about 50% of its
sequence conserved between E. coli and human, and some domains are 96% similar. Second,
several of the major heat shock proteins are members of gene (protein) families that include
proteins normally present and, in most cases, essential for cell function.
The hsp70 of yeast is the most thoroughly studied heat shock family system; it consists of nine
members that differ to varying degrees in sequence and in cellular localization. Three members
are localized to specific compartments of the cell: the mitochondrion, the nucleus, and the
endoplasmic reticulum. They differ also with regard to the conditions under which they are
synthesized. In fact, it is the discovery of the function of hsp70 proteins made in normal cells
that provided the data for the chaperone model.

Events in the Heat-shocked Cell

The suggestion that, one of the major effects of heat shock is an unfolding, or an incomplete or
improper folding of proteins, appeared early in the heat shock literature. Many recent studies
have confirmed this hypothesis (Ref. Nguyen, V. T., Morange, M., and Bensaude, 0. (1989) J.
Biol. Chem. 264) and, clearly, the concept that heat shock proteins function as chaperones
provides further strong support for these ideas. An interesting extrapolation of this hypothesis
is that it is the improperly folded polypeptide that actually initiates the induction of the heat
shock response. In bacteria, manipulations of protein structure that lead to increased amounts
of badly folded proteins turn on the heat shock regulon. In animal cells, there is a temperaturesensitive mutant with a defect in the ubiquitin-dependent proteolytic degradation pathway that
leads to accumulation of defective proteins. This mutant shows enhanced levels of heat shock
proteins under non-heat shock conditions. In contrast, the addition of deuterium oxide or
glycerol, agents that can stabilize proteins, inhibits and delays the induction of heat shock
proteins.
In addition to improper polypeptide folding, heat shock leads to a plethora of changes that are
dependent on both the intensity of the stress and on the cell system. These include effects on
macromolecular synthesis, on levels of cations, on states of protein phosphorylation, on
metabolic pathways, on cytoskeleton networks, etc.
A number of other proteins have been reported to be heat shock proteins, based on their
enhanced rate of synthesis after stress. In E. coli, the heat shock regulon includes about 20
genes, and there is probably twice that number in the eukaryotic cell. Some of these have been
identified as glycolytic enzymes, i.e. glyceraldehyde-3-phosphate dehydrogenase, enolase,
phosphoglycerate kinase, and others are noted by activities such as a collagen-binding protein
and heme oxygenase. We can rationalize the appearance of the former set since stressed cells
tend to shift from aerobic to glycolytic metabolism for energy production. But others remain
enigmatic. The most predominant in this latter category is a set of small molecular weight stress
proteins that vaguely resemble each other in hydropathy profiles but not sequence, except for
a short region that is homologous to a sequence in the mammalian lens (Y crystallin). These low
molecular mass stress proteins (15-30 kDa) are most abundant in stressed plants where some
are found as RNA nucleoproteins and others are translocated to the chloroplasts and to the
mitochondria. In all eukaryotic cells, they can form huge granular arrays. Like some of the heat
shock proteins noted above, these small proteins are found during various stages of embryonic
development and, in yeast, during sporulation.

Why Are Heat Shock Proteins Important?

The function of a protein is determined by its three-dimensional structure. When excessive
heat is applied to proteins, chains of amino acids which are folded into spirals, loops and sheets
begin to lose their shapes. When the interior of these proteins gets exposed, proteins can
adhere and form globs. This can make them dysfunctional. Protein conformational defects are
responsible for a number of pathologies, ranging from Alzheimer's disease and oncogenic
transformation in humans to heat and drought susceptibility in plants. Chaperones protect
against denaturization. Heat Shock Proteins bind to denatured proteins to prevent
aggregation. Some Heat Shock Proteins, like Hsp104, have the ability to rescue already
aggregated proteins.

Why Don't Heat Shock Proteins Denature?

-Better Hydrogen Bonds
-Better Hydrophobic Internal Packing
-Enhanced Secondary Structure
-Helix Dipole Stabilization

Definitions:
Chaperone: In molecular biology, molecular chaperones are proteins that assist the noncovalent folding or unfolding and the assembly or disassembly of other macromolecular
structures.
Upregulation and downregulation: Downregulation is the process by which a cell decreases the
quantity of a cellular component, such as RNA or protein, in response to an external variable.
An increase of a cellular component is called upregulation.
Hsp 90: The Hsp 90 kilodalton alpha (HSP 90-alpha) is a protein that in humans is encoded by
the HSP90AA1 gene.
Hsp 70: The 70 kilodalton heat shock proteins (Hsp70s) are a family of conserved ubiquitously
expressed heat shock proteins. Proteins with similar structure exist in virtually all living
organisms. The Hsp70s are an important part of the cell's machinery for protein folding, and
help to protect cells from stress.

References:
1. Heat Shock Proteins
Milton J. Schlesinger
From the Department of Molecular Microbiology,
Washington University School of Medicine,
St. Louis, Missouri 63110
Link:
http://mityeast.pbworks.com/w/file/fetch/55833870/minireview%20Heat%20Shock%20
Proteins.pdf
2. Wikipedia
3. Link:http://www.cs.stedwards.edu/chem/Chemistry/CHEM43/CHEM43/HSP/STRUCTUR
E.HTML