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Insulin: Weight = 5733, 51 amino acids Glutamine Synthetase: Weight = 600,000, 468 amino acids Amino Acids & Dehydration Synthesis Amino acid structure: Functional groups Peptide bonds Sample amino acid structures: carboxyl group-blue alpha carbon-gray amino group-green R groups-beige The twenty essential amino acids. Note R groups in blue Amino Acids w/ various R groups Nonpolar amino acids Neutral Amino Acids Ionic amino acids Amino acids are joined together in proteins by peptide bonds. • A peptide bond forms between the carboxyl group of one amino acid (amino acid 1 in the figure preceding) and the amino group of the adjacent amino acid (amino acid 2). Dehydration synthesis Dehydration synthesis animation Dehydration Synthesis Dehydration Synthesis: Amino acid 1 Amino acid 2 Amino group Dipeptide (Peptide bond) Protein Structure • Amino acid structure • Dehydration synthesis Insulin: Weight = 5733, 51 amino acids Glutamine Synthetase: Weight = 600,000, 468 amino acids Four ‘levels’ of protein structure Four levels of protein structure Primary structure: Visualized as a straight chain of aa’s, but with a specific sequence, As determined by its gene Secondary structure: alpha helix or beta pleated sheet Tertiary structure: coiled chains of aa’s are folded & wound around themselves Close up of 2o & 3o Protein Structure Quarternary structure: separate polypeptide chains are combined Levels of Protein Structure Four Levels of Protein Structure (note change in scale from 2o to 3o) Collagen molecule Actin molecule Myosin molecule Hemoglobin molecule Antibody molecule (purple) Reverse transcriptase of HIV1 w/ a fragment of DNA (colors) Protein Synthesis: Transcription & Translation Transcription: Making a copy of the blueprint Morse Code Key Using the Genetic Code: DNA :: mRNA :: Protein • Get from ‘language’ of DNA (A,G,C,orT) to ‘language’ of protein (aa’s) • DNA’s ‘language’ is a triplet code in which 3 nucleotide bases (a codon) specify 1 amino acid in protein. DNA (1 gene) :: mRNA :: polypeptide Steps in Transcription Transcription: Note the free nucleotides DNA unzips Complementary base pairing mRNA- final product of Transcription mRNA moves off to ribosome Transcription: Getting the plan straight (copying the gene) Transcription animation Transcription animation Translation Building the protein from the plan Using the Genetic Code: DNA :: mRNA :: Protein • Get from ‘language’ of DNA (A,G,C,orT) to ‘language’ of protein (aa’s) • DNA’s ‘language’ is a triplet code in which 3 nucleotide bases (a codon) specify 1 amino acid in protein. Structure of a Ribosome Structure of a tRNA Translation: purple = mRNA blue = ribosome yellow = tRNA (note anticodon) white = amino acids red = peptide bond Translation initiation Translation initiation (cont) Translation- elongation Translation- termination A Polysome: With more than one ribosome translating an mRNA at one time, it is possible to produce many polypeptides simultaneously from a single mRNA. Protein Synthesis In a prokaryotic cell, transcription and translation are coupled; that is, translation begins while the mRNA is still being synthesized. In a eukaryotic cell, transcription occurs in the nucleus, and translation occurs in the cytoplasm. Protein synthesis in eukaryotes Antibiotic Mechanism Antibiotic Mechanism Collagen molecule Actin molecule Myosin molecule Antibody molecule Hemoglobin molecule Enzymes Definitions: • Catalyst = An additive that speeds up a chemical reaction without itself being consumed or changed by the reaction • Enzyme = A protein that acts as a catalyst, usually in a biological context. – All enzymes are proteins, not all proteins are enzymes Enzyme mechanism of action: An enzyme improves the odds of ‘useful’ collisions between substrate molecules. Steps in enzyme function Enzyme-substrate animation S = substrate E = enzyme P = product ES = enzymesubstrate complex Lock & Key Model: Enzymes are very specific and it was suggested by Emil Fischer in 1890 that this was because the enzyme had a particular shape into which the substrate(s) fit exactly. This is often referred to as "the lock and key" hypothesis. An enzyme combines with its substrate(s) to form a shortlived enzyme-substrate complex. Induced fit hypothesis: In 1958 Daniel Koshland suggested a modification to the "lock and key" hypothesis. Enzymes are rather flexible structures. The active site of an enzyme could be modified as the substrate interacts with the enzyme. The amino acids side chains which make up the active site are molded into a precise shape which enables the enzyme to perform its catalytic function. In some cases the substrate molecule changes shape slightly as it enters the active site. A suitable analogy would be that of a hand changing the shape of a glove as the glove is put on. Enzyme Animation Enzyme Websites: • Enzyme notes in ppt format, few diagrams: • http://www.hcc.mnscu.edu/programs/dept/chem/V.28/page_id_2897.html • animation of enzyme action • http://web.ukonline.co.uk/webwise/spinneret/other/hienz.htm • written outline- enzymes by Worthington • http://www.worthington-biochem.com/introBiochem/introEnzymes.html • asd Enzyme & Cell Regulation • Gene activation • Feedback loops for enzyme activity Various possible routes of feedback in the production-regulation of a given protein Changing the conformation (shape) of an enzyme’s active site changes its ability to act as a catalyst Zymogens: Enzyme Precursors Competitive Enzyme Inhibition Allosteric Modulation of an Enzyme Genes can be either inducible or repressible. Many genes are normally blocked by the action of a repressor protein. This prevents the RNA polymerase enzyme from binding to the gene and transcribing the structural gene. Such genes are induced by the arrival of an inducer molecule which binds to the repressor protein and rendering it inactive. This allows transcription from the structural gene and the production of a protein. Other genes are normally active and able to be constantly transcribed, because the repressor protein is produced in an inactive form. On the arrival and binding of the corepressor molecule the complex can act as a functional repressor and block the structural gene by binding at the operator site. Steps in Transcription DNA Technology Bases: A- adenine G- guanine T- thymine C- cytosine Other translation animations: • Add an amino acid to tRNA• http://www.phschool.com/science/biology_place/biocoach/translation/addaa.html • Initiation of Translation• http://www.phschool.com/science/biology_place/biocoach/translation/init.html • Elongation of Polypeptide• http://www.phschool.com/science/biology_place/biocoach/translation/elong1.html • Termination of Polypeptide• http://www.phschool.com/science/biology_place/biocoach/translation/term.html