Post-translational Modifications to Proteins

Affecting Form and Function

 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


modification site may be a binding interface

 Subcellular


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
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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


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

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

proteins against proteolysis

Limit approach of macromolecules to protein surface


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

sorting signals

M6P for lysosomal hydrolases  GPI anchors (see later)

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

of core protein and polysaccharide chains
Extremely diverse

 

hydrated gel

Resists compressive forces Regulate traffic (perlecan in kidney)


regulate secreted protein activity
e.g. chemokines in inflammatory response

Membrane proteins: GPI anchors


proteins are delivered to the apical plasma membrane Trypanosomes can shed these proteins to avoid immune attack

Myristylation and Farnesylation

cytosolic proteins to the plasma membrane Protein usually involved in signal transduction

Proteolytic processing


is this common for secreted enzymes?
Some peptides (e.g. enkephalins) too short by themselves  Prevent premature activation of hydrolytic enzymes

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
 Cell

signals from cell surface

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

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 and Histones
 Acetylation

enhances transcription  Deacetylation represses transcription

Small Molecule Binding

Retinal B. Heme group

Protein Degradation
 Degradation
 Ubiquitin

ligase  Degradation signal

 Multiubiquitin

chain marks protein for degradation in proteosome

Common Post-translational Modifications
Sulphydryls Amines Disulphide bond Cysteinylation Methylation Acetylation Farnesylation Biotinylation Stearoylation Pyroglutamic acid Carboxylation Phosphorylation Pentoses Hexosamines N-acetylhexosamines Oxidation Glutathionylation Formylation Lipoic acid Myristoylation Palmitoylation Geranylgeranylation Deamidation Sulphation Deoxyhexoses Hexoses Sialic acid

Acids & amides Hydroxyl groups Carbohydrates

 Post-translational
 Correct

modifications – necessary for protein function
protein folding  Organism development  Cellular Signalling  Motor Proteins  Regulating degradation  …and much more…