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Index >> Cellular Components >> Protein Synthesis

Protein Synthesis

Protein Synthesis- The process of peptide synthesis translation of m-RNA into peptides, can be divided into three stages-initiation, elongation and termination. Thanks to the development of a variety of techniques, each one of these stages can now be studied as partial reactions of the overall process of translation.

The process of initiation of polypeptide synthesis needs, in addition to mRNA and formylmethionyl t-RNA, the ribosomes, GTP and at least three initiation factors, IF1, IF2, and IF3. IF1 and IF3 are involved in the binding 30 S ribosomal particle to m-RNA. IF2 is involved in the recognition of correct codon of m-RNA by formyl-methionine t-RNA. GTP is involved in the positioning of formyl methionine t-RNA and is subsequently hydrolysed to GDP and Pi. Methionine plays a very important role in peptide initiation in E. coli.

However, it is incorporated as N-formylmethionine as the first amino acid in the peptide chain. The formylation occurs after the attachment of the methionine to the t-RNA. Formylation of the amino group in methionine prevents peptide bond formation with the free carboxyl group of another amino acid but mimics a peptide bond which is recognized by a ribosomal site.

The process of peptide chain elongation involves binding of amino acyl-t-RNA to ribosomes, translocation to another ribosomal site of the growing peptide and the formation of the peptide bond. These three stages enable correct reading of the information from the m-RNA. The key step in the correct translation of m-RNA, is the specific interaction between the anticodon of the t-RNA and the codon in the m-RNA, which ensures that the correct amino acid is lined up for peptide bond formation.

There are at least two proteins involved in peptide chain elongation designated as, EF1 and EF2 which are involved in the formation of GTP complexes and subsequently modified with the hydrolysis of GTP to GDP. These factors mediate the bond formation between the free carboxyl of the formyl methionine with the amino acid adjacent to it.

This step is catalysed by the enzyme peptidyl transferase. In subsequent reactions peptide bond is formed between the terminal carboxyl of the growing polypeptide and a amino group of the amino acyl-t­RNA. This reaction needs Mg ++, K+ or NH4+ ions. GTP is involved in the process of transferring peptidyl-tRNA from one site to another on the ribosome, displacing the free t-RNA. The elongation factor EF2 catalyses this step in which the ribosome is also translocated to read a new codon in the m-RNA.

Chain elongation continues through aminoacyl-t-RNA binding, peptide bond formation and translocation until the ribosomes reach the termination codons (UAG, UAA or UGA) on the m­RNA. The peptide chain is released at this point and this is mediated by at least two protein release factors RFI and R F2. The ribosome m-RNA complex at this stage dissociates and dissociation factor RF3 appears to be responsible in dissociating 70 S ribosome into 30 S and 50 S units. The subunits are then again ready to from complex with a new m-RNA and initiate synthesis of an another peptide.

 

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