Lecture4 Synthesis of Proteins. Lecture4 Synthesis of Proteins.

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Lecture4 Synthesis of Proteins

The Three Roles of RNA in Translation Messenger RNA (mRNA) Transfer RNA (tRNA) Ribosomal RNA (rRNA)

Universal Genetic Code Degeneracy: a particular amino acid can be specified by multiple codons. Codon usage bias: refers to differences in the frequency of occurrence of synonymous codons in coding DNA

The Folded Structure of tRNA Promotes Its Decoding Functions Some 30-40 different tRNAs have been identified in bacterial cells and as many as 50-100 in animal and plant cells.

aminoacylation of tRNAs The conservation of the genetic code suggests that aminoacyl-tRNA synthetases evolved early and were possibly among the first protein enzymes to emerge from the RNA world

Nonstandard base pairing at the wobble position

Initiation Selection of the initiating AUG is facilitated by specific surrounding nucleotides called the Kozak sequence, (5') ACCAUGG (3').

Elongation

Termination

Polysomes and Rapid Ribosome Recycling Increase the Efficiency of Translation

Purifying, Detecting, and Characterizing Proteins A protein must be purified before its structure and the mechanism of its action can be studied. The two most widely used characteristics for separating proteins are size, defined as either length or mass, and binding affinity for specific ligands.

Centrifugation Proteins vary greatly in mass but not in density. Unless a protein has an attached lipid or carbohydrate, its density will not vary by more than 15 percent from 1.37 g/cm3, the average protein density. Differential Centrifugation Rate-Zonal Centrifugation Isopycnic centrifugation

SDS-polyacrylamide gel electrophoresis (SDS-PAGE)

Two-Dimensional Gel Electrophoresis