Translation III: The mechanism of protein synthesis

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Translation III: The mechanism of protein synthesis Edited by Gergely Berta

The steps of initiation Initiation factors B P-site (peptidyl-tRNA binding site) A-site (aminoacyl-tRNA binding site) C 50S fMet formylmethionyl-tRNA 30S anticodon 2 codons 5’ A U G mRNA Ribosome binding sequence

N-formylmethionyl-tRNA formyl group C HN-CH-C O O R1 A O Blocked amino acid → it can be used only at the beginning, i.e. at the N-terminus The formyl group is removed by deformylase during elongation Aminopeptidase removes methionine Eukaryotes: no formylation

Elongation of the translation E-site (exit-site) B P-site (peptidyl-tRNA binding site) A-site (aminoacyl-tRNA binding site) C fMet Aa2 EF 5’

B.: formation of the peptide bond P-site A-site Peptidyl transferase cuts (hydrolyzes) the ester bond between the amino acid and the tRNA in the P-site Energy is released The carboxyl group of the amino acid in the P-site can now bind to the amino group of the amino acid in the A-site The peptide bond is created O H H O O O C HN-CH-C C HN-CH-C N-CH-C H2N-CH-C O O O H O O R1 H R1 R2 R2 A A A O O O

Termination of the translation aan aan-1 aan-2 aan-3 aan-4 aan-5 aan-6 aan-7 aan-8 aan-9 releasing factor stop codon

3D structure http://publications.nigms.nih.gov www_wadsworth.org_joachim_frank.jpg

Eukaryotic translation Localisation of the ribosomes: Free ribosomes in the cytoplasm, they synthetise: Proteins of the cytosol Proteins of the nucleus Proteins of the mitochondria Bound ribosomes on the surface of the rough endoplasmic reticulum: Secretory proteins Proteins of the rough ER Proteins of the Golgi-apparatus Proteins of the lysosomes Proteins of the cell membrane

General features of translation Direction: 5’→3’ (mRNA) N-Terminus → C-Terminus (polypeptide chain) Polysomes are formed one polypeptide chain/ ribosome The 3D structure of the protein chain is being already formed during translation

Inhibitors of protein synthesis In prokaryotes: Chloramphenicol peptidyl transferase Erythromycin translocation Tetracyclin aminoacyl-tRNA binding Streptomycin 30S subunit What is the relevance? These are used as antibiotics against bacterial infections They have side-effects: Mitochondrial translation is also inhibited If they are not specific enough for prokaryotes They influence other proteins/mechanisms in the eukaryotic cells