Post-translational

Modifications to Proteins
Affecting Form and Function
Contents
 Why are proteins modified?
 Glycosylation
 Membrane proteins
 Proteolytic processing
 Phosphorylation
 Acetylation
 Small Molecule Binding
 Regulated degradation
Why are proteins modified?
 Regulation of activity
 modification may turn activity on
 modification may turn activity off
 modification may generate a different function
 Protein-protein interaction
 modification site may be a binding interface
 Subcellular localization
 modification site may be a targeting signal
 modification may be a membrane anchor
 Aging
 modification may identify the protein for degradation
 modification may target a protein to be scavenged
Topics to cover in my section
 593-595 Membrane proteins - myristlyation, farnesylation

 598,612-3,735-736 Glycosylation
 702-704 Glycosylation in rough ER
 706,763-764 GPI anchors
 733-735 Oligosaccharide processing (Golgi & ER, Glyco-related)
 742-745 M6P in lysosomal enzymes
 1085-6 Lectins and the immune system
 1091-6 Proteoglycans and extra-cellular matrix

 678-681,685,694 Cleavage of signal sequences (mitochondria, chloroplasts, ER)
 760 Proteolysis of secreted proteins (to activate them)
 893-895 Notch cleavage in neuron development
Glycosylation
Major form of protein modification
Sugars are added in the ER and Golgi
Most proteins formed in the ER are
glycoproteins
Many different forms and functions
Initial glycosylation in the ER

A precursor
oligosaccharide
is formed on a
dolichol lipid
This is
transferred to
the growing
protein
Glycosylation and protein folding

by glucosidase
Processing in the Golgi
Functions of glycosylation
Stabilise proteins against proteolysis
 Limit approach of macromolecules to protein
surface
Modulation of immune response
 Selectins (weakly) bind to oligosaccharides
 Helps to concentrate lymphocytes in lymphoid
organs
 Attracts white blood cells & platelets to
inflammation sites
Functions of glycosylation
Provide sorting signals
 M6P for lysosomal hydrolases
 GPI anchors (see later)

Contributes to differentiation events in

organism development
 Removing N-acetylglucoasmine transferase I in
mice causes embryo death
 Neural
tube development and left-right body plan
asymmetry impaired
Proteoglycans and the Extracellular
Matrix
Made of core protein and
polysaccharide chains
 Extremely diverse
Form hydrated gel
 Resists compressive forces
 Regulate traffic (perlecan in kidney)
Can regulate secreted
protein activity
 e.g. chemokines in inflammatory
response
Membrane proteins: GPI anchors

GPI-anchored proteins are delivered to

the apical plasma membrane
Trypanosomes can shed these proteins
to avoid immune attack
Myristylation and Farnesylation
Attaches
cytosolic
proteins to
the plasma
membrane
Protein
usually
involved in
signal
transduction
Proteolytic processing

Why is this common for secreted

enzymes?
 Some peptides (e.g. enkephalins) too short
by themselves
 Prevent premature activation of hydrolytic
enzymes
Phosphorylation
 Most common
posttranslational modification
to proteins in eukaryotes
 Enzymes and regulators are
turned ‘on’ and ‘off’
 Energy from ATP
Phosphorylation Regulates Protein
Synthesis – eIF-2
Phosphorylation and Molecular
Switches
Signalling using GTP-Binding - Ras
Protein
 Broadcasts signals from cell surface
 Cell proliferation
 Differentiation
Phosphorylation and Motor Proteins
 Move chromosomes during
mitosis
 Move organelles along
molecular tracks
 Move enzymes along DNA
during DNA synthesis
Phosphorylation and Motor Proteins
 ATP binding - conformation 1
to conformation 2
 ATP hydrolyzed to ADP Pi -
conformation 2 to
conformation 3.
 Release of ADP and Pi -
back to conformation 1.
 Irreversible – one direction
only
Acetylation
Acetylation and Histones
 Acetylation enhances
transcription
 Deacetylation
represses transcription
Small Molecule Binding

A. Retinal
B. Heme group
Protein Degradation
 Degradation
mechanisms:
 Ubiquitin
ligase
 Degradation signal

 Multiubiquitin
chain marks
protein for
degradation in
proteosome
Common Post-translational Modifications

Sulphydryls Disulphide bond Oxidation

Cysteinylation Glutathionylation
Amines Methylation Formylation
Acetylation Lipoic acid
Farnesylation Myristoylation
Biotinylation Palmitoylation
Stearoylation Geranylgeranylation
Acids & amides Pyroglutamic acid Deamidation
Carboxylation
Hydroxyl groups Phosphorylation Sulphation
Carbohydrates Pentoses Deoxyhexoses
Hexosamines Hexoses
N-acetylhexosamines Sialic acid
Summary
 Post-translational
modifications –
necessary for protein function
 Correct protein folding
 Organism development

 Cellular Signalling

 Motor Proteins

 Regulating degradation

 …and much more…