Understanding functional
architechture gives us POWER
to:
Diagnose and find reasons for
diseases
Create modifying drugs
Engineer our own designer-
proteins
Protein structure determines function
DNA
(mRNA)
Translation:
Modifications:
Chemical modification of aminoacids
Interaction with other molecules
Proteolytic cleavage
(Location)
New 3D structure
3D structure determines function:
New function
a Proteins are single, unbranched chains of amino acid monomers
a There are 20 different amino acids
a The amino acid sidechains in a peptide can become modified, extending the functional repetoire of aminoacids to more
than hundred different amino acids.
a A proteins amino acid sequence determines its three-dimensional structure (conformation)
a In turn, a proteins structure determines the function of that protein
a Conformation (=function) is dynamically regulated in several different ways
All amino acids have the same general structure
but the side chain (R group) of each is different
R: Hydrophilic:
Basic
Acidic
Non-charged
Hydrophobic
Special
Hydrophilic amino acids
Hydrophobic
and special amino acids
Peptide bonds connect amino acids into linear
chains
Backbone
Side-chains
Side chain modifications change the chemical
(functional) properties of proteins
Acetylation
Phosphorylation
Hydroxylation
Methylation
=> Expanding the
repetoire of existing
amino acid side-chains to
Carboxylation > 100 variations!
Glycosylation
Ubiquitylation
Four levels of structure determine the shape of proteins
-strand -helix
Secondary structure
Sheet
Helix
(U)-turn
Tertiary structure
Tertiary structure
Different graphical representations of the same protein
(tertiary structure)
Quaternary structure
Multiprotein complexes: molecular machines
Sequence homology suggests functional and
evolutionary relationships between proteins
When the stucture of a newly discovered protein is known, comparison to other proteins
across species can help predict function
Folding, modification, and degradation of proteins
An amyloid plaque in
Alzheimers disease is
a tangle of protein
filaments
The information for protein folding is encoded in the
sequence
Folding of proteins in vivo is promoted by chaperones
Large proteins with a lot of secondary structure may require assisted folding to avoid
aggregation of unfolded protein
- Molecular chaperones and chaperonins prevent aggregation of unfolded protein
Folding of proteins in vivo is promoted by chaperones
Large proteins with a lot of secondary structure may require assisted folding to avoid
aggregation of unfolded protein
- Chaperones and chaperonins prevent aggregation of unfolded protein
Functional design of proteins
a Allosteric transitions
` Release of catalytic subunits, active inactive states, cooperative binding
of ligands
a Chemical modification:
` Phosphorylation, acetylation etc. dephosphorylation, deacetylation etc.
a Proteolytic activation
a Compartmentalization
Protein degradation via the ubiquitin-mediated pathway
ATP
Cooperative binding of
calcium: binding of one
calcium enhances the
affinity for the next
calcium
When 4 calcium are
bound a major allosteric
conformational change
occurs
Calmodulin is a switch
protein because this
effect in turn regulates
other proteins bound by
the compact calmodulin
Another class of switch proteins: GTPases
Chemical modification
a Allosteric transitions
` Release of catalytic subunits, active inactive states, cooperative binding
of ligands
a Chemical modification:
` Phosphorylation, acetylation etc. dephosphorylation, deacetylation etc.
a Proteolytic activation
a Compartmentalization
Example containing all levels of regulatin of protein activity
GFP-tagged GLUT4
Now that you KNOW the basic principles of protein structure and
function you can UNDERSTAND:
SDS-polyacrylamide
gel electrophoresis
Stationary phase:
Separation of proteins by charge: ion exchange
chromatography
Matrix-Assisted-Laser-Desorption/Ionization Time-of-
flight mass spectrometry (MALDI-TOF MS)
MS spectrum
Example of a proteome analysis workflow
Cell/tissue of interest
Isolate organelles
(fractionation)
Confirm organelle-specific
proteins
Subfractionate, detect
peptides, identify
corresponding proteins
X-ray crystallography is used to determine protein
structure