Post-translational modifications of proteins : Acetylcholinesterase as a model system

Contents
1. 2. 3. 4. 5. 6. 7. 8. Introduction Amino acid modification Bacterial toxins and post-translational modifications Glycosylation Other lipid modifications Protein turnover Limited proteolysis and membrane protein secretases Proteolysis of the amyloid precursor protein(APP) and Alzheimers disease 9. Post- translational modification of Ache : its physiological role and significance

1. Introduction

2. Amino acid modifications

# N-terminal protein modifications

Removal of N-terminal methionyl residue (by methionine amino-peptidase ) : occurs when second residue is one of the smallest amino acid
correlate with stability, turnover of protein, function

N-termial methionine Cleavage (bacteria yeast gene deletion ) Acetylation Methyation Phosphorylation

# C-terminal protein modification

: less diverse than N-terminal but include amidation and glycosyl-phosphatidylinositol(GPI) anchor addition

# Nonenzymic modification ex) vasodilator (NO) : haemoglobin myoglobin cystein residue binding

3. Bacterial toxins and post-translational modifications

Ex) cholera and pertussis toxins : modify G proteins through ADP-robosylation

4. Glycosylation N-linked, O-linked, GPI-linked glycosylation is not normally a reversible phenomenon and does not play a role in regulation of activity

: maintain the solubility of glycoprotein ensure the correct folding of the extracellular domains and stability of the protein against degradation protein targeting and recognition

5. Other lipid modifications Myristoylation, palmitoylation, prenylation Ex) addition of myristic acid : N-term sequence Met-Gly after removing of the initiation Met myristate is attached to the glycine residue Many signalling protein are N-myristoylatied : many protein kinase and phosphatases, various G proteins, Ca-binding protein protein etc..

6. Protein turnover
Degradation of many cellular protein requires the Covalent addition of ubiquitin as a signal for proteolysis By the proteasome Post-translaional modification : addition of multiple ubiquitin

ubiquitin-activating enzyme ubiquitin-conjugating enzyme ubiquitin-protein ligase Many regulatory proteins are degraded by the ubiquitin system

7. Limited proteolysis and membrane protein secretases

Limited proteolysis of cell-surface protein are cleaved By the membrane protein secretase like ACE secretase

8. Proteolysis of the amyloid precursor protein (APP) and Alzheimers disease

1) Intracellular neurofibrillary tangles 2) Extracellular senile plaques abnormal post-translational modification

tau protein : abnormal hyper-phosphorylated form in the case of the tangles O-GlcNAc-modified in normal modification

9. Post-translational modification of AChE its physiological role and significance In different species of animals and in various tissues : conserved catalytic domain + variable C-terminal domain (by alternative splicing) Different molecular forms of AChE are characterised by different localisation Soluble form : plasma, cerebrospinal fluid, venom of snakes GPI-anchored form : surface of blood cells Collagen-tailed form : neuromuscular junction

10. Concluding remarks Knowing the sequence of the human genome is Only a first step towards understanding the complexity Of the protein component expressed in cell Technologies must continue to be developed to Allow analysis of the these modification