Chem 1100 Lecture 25Outline of Last Lecture I. BiologyII. CellsIII. DNAOutline of Current Lecture I. ProteinsII. From DNA to ProteinIII. Lab Synthesis of ProteinsIV. Mixing GenesCurrent LectureI. Proteinsa. Amino acid polymers – 20 R groupsb. Process repeats over and over, creating a peptide chain (short protein)c. Amino acid residues: amino acids that are incorporate into a peptide/polymerd. Each amino acid has to be in the right order for the protein to function properlye. Codons: 3 DNA bases to code for 1 amino acid in a proteinf. How are codons relevant in genetic expression?  The order of DNA bases  order of amino acids in a proteinII. From DNA to Proteina. DNA  TAC AAA TTT ATTb. RNA  AUG UUU AAA UAAc. Protein  Met Phe Lys d. Protein Sequencei. Order of amino acids in the polymerii. Determines shape, reactivity, active site location, etc.iii. NH2 group on one end (N-terminus)iv. COOH group on the other (C-terminus)v. Start with N and go to Cvi. Primary structure: linear sequence of amino acids and the location of any disulfide (-S-S-) bridges.vii. Second structure: folding pattern within a segment of the protein chain.viii. Tertiary structure: 3D folding of the entire molecule. Sequence  amino terminal (N-terminal) at one end carboxyl terminal (C-terminal) at the other. These notes represent a detailed interpretation of the professor’s lecture. GradeBuddy is best used as a supplement to your own notes, not as a substitute.e. Putting it Togetheri. 3D structure brings amino acids togetherii. Determines the arrangementiii. Determines the active site (catalyzes reaction)iv. Not all proteins have an active sitev. Enzymes have an active site  a place where the starting materials can bind. Enzyme is not used up it’s just ready to bind since its catalytic.f. Active Sitei. Enzymes will bind the substrates and generate products. ii. Promotes bond making or bond breakingiii. When chains of amino acids fold they form the most stable structure that they can. If there are problems with the primary structure there will probably be problems with the other structures as well. iv. Sickle cell anemia – exists in hemoglobin. 2 amino acids change in the hemoglobin when sickle cell anemia occurs. 574 total amino acids in hemoglobin and only 2 of those get chained. Sickle celled shaped RBCs can’t get through the capillaries. They also can’t get through the organs. Small changes equal big problems with amino acids. g. Changing Amino Acidsi. Different DNA sequence  different amino acidsii. Hereditary Mutation: if you have sickle celled anemia you can pass that on through your geneticsiii. Normally nature will weed out bad cells. But the reason sickle celled anemia is still around is they think that maybe it protects against malaria so the body doesn’t fight against it that hard. III. Lab Synthesis of Proteinsa. Hemoglobin can not yet be synthesizedi. People aren’t even working on it right now.ii. Ribonuclease A has been synthesized. It folds into an active protein. iii. But there’s a better way..iv. Through Recombinant DNA Technology.b. Recombinant DNAi. Take a gene and put it into a bacterium and that bacterium is going to make the protein. ii. Bacteria grow very quickly, that is why they are preferred. iii. Toad protein into E.coli  E.coli can make the toad protein very quicklyiv. It’s active because it’s a biological system and the active protein is there.c. Insulini. Enzymeii. Regulates blood glucoseiii. Diabetes = low insulin productioniv. High blood glucose = sticky bloodv. We used to get insulin from slaughtered pigs and cows. It was an expensive process to isolate the protein. Also it’s different than human insulin. Not as effective.vi. Insulin can now be made in the laboratory but its’ very expensive and very complicated and doesn’t always come out right.vii. So that’s when we started using bacteria to do the work for us.viii. Plasmids: A plasmid is a small DNA molecule within a cell that is physicallyseparated from a chromosomal DNA and can replicate independently.ix. Never been found in an eukaryotes.x. Plasmid DNA: It’s a small ring and easy to isolate. They have restriction enzymes that will search for a certain base sequence and it will cut at thatcertain sequence. Allows insertion of “alien” genes. xi. Vector: a plasmid that has the alien DNA in itxii. Clone: cells that are derived from the same parent cell (genetically identical)xiii. Gene therapy: supplying cells with normal genes so they can start reproducing insulin themselves instead of having to give the human directinjection of insulin whenever they are low. d. Harvest genes isolate plasmidse. Harvested protein isolates the proteins that are madef. Clinical Usei. HGH : human growth hormoneii. HGH-deficient  can lead to dwarfism. Inject HGH when you are young and can combat dwarfismiii. The old way was to get it from human cadaversiv. The new way is to get it from bacteria which both ways are still expensiveIV. Mixing Genesa. Insert DNA into a plant or animal and it will change the genes of their offspringb. Transgenic Speciesc. Change the genes and you’ll change the proteins the cell makesd. The cell reproduces and organism with new characteristics is formed from the different DNA.e. Crops: traditional way of doing it is cross breedingf. New way they just insert the genes straight into the DNA materialg. Results of Mixing Genes:i. Herbicide Resistanceii. Resist Diseasesiii. Improved Nutritioniv. Temperature Resistancev. Better Yield of Cropsvi. Production of Pharmaceuticalsh. The Futurei. Cows that drink Human milk ii. Animals grow fasteriii. Low fat meatsiv. Vaccine productionv. Plastic Productioni. Molecular Pharmingi. Induce animal cells to make medsii. Pigs produce human hemoglobiniii. Insuliniv. Hepatitis C Vaccinev. Concerns? Environmental, monarch butterfly, allergic reactionsj. Cloning Mammalsi. Genetically identical offspringii. Make this animal to create a specific proteiniii. Engineer animals that make a desired substanceiv. Want more engineered animals  productionv. If you breed more you can lose the genevi. Cloning = no gene lostvii. Cystic Fibrosis treatment: Elastase inhibitor  needs improvementk. Stem Cellsi. Undifferentiated cellsii. Idea: stem cell  specialized celliii. Embryonic stem cells  come from embryosiv. Other sources  umbilical cord and you can get some from adults as