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Download Name: Cell Biology Test #1: 50 points
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Unit Written Questions: 10 points Please write your answers on the paper provided: Abbreviations for the 20 different amino acids: Phenylalanine-Phe, Leucine-Leu, Isoleucine-Ile, MethionineMet, Valine-Val, Serine-Ser, Proline-Pro, Threonine-Thr, Alanine-Ala, Tyrosine-Tyr, Histidine-His, GlutamineGin, Asparagine-Asn, Lysine-Lys, Aspartate-Asp, Glutamate-Glu, Cysteine-Cys, Tryptophan-Tyr, ArginineArg, and Glycine-Gly. --------------------------------------------------------------------------------------------------------------------59) 5 pt: Examine the mRNA below and determine the protein (aa sequence) that would result from successful translation by a ribosome. a) # of amino acids ______ b) # of codons?_____ c) # of peptide bonds in protein?_____ d) Find the start codon and show the specific amino acids (names) in the specific sequence of the peptide resulting from translation of this mRNA. 5’-AGCCUUAUGAGGCAGGGGAUGACCGAGCGCUAAGGCCGG-3’ Methionine-Typtophan-Glutamine-Glycine-Methionine-Threonine-Stop codon Seven codons (last is stop codon-not an amino acid) Six amino acids Five peptide bonds 60) 2pt: The mRNA above was transcribed from a single template strand of DNA that was unwound from a double helix. What was the base sequence of this template ssDNA? 5’-AGCCUUAUGAGGCAGGGGAUGACCGAGCGCUAAGGCCGG-3’ 3’-TCGGAATACTCCGTCCCCTACTGGCTCGCGATTCCGGCC-5’ ssDNA 61) 3 pt: Compare and contrast the function of the smooth endoplasmic reticulum, the rough endoplasmic reticulum and free ribosomes that are not bound to the ER? 20-40 words rRNA: smaller segments of RNA that make up the large and small subunit of a ribosome. These two subunits permit translation of mRNA to a peptide sequence and create docking sites for tRNA. tRNA: 64=61 different types of tRNA can deliver amino acids to the A-site of a ribosome, the anticodon of the tRNA must match (antiparallel) the codon of the mRNA mRNA: this caries the codons (including start-methionine and stop-no tRNA) that are translated (3 amino acids at a time) into an amino acid sequence micoRNA: These are the RNA fragments that help fine tune transcription and probably even translation of our genetic information into a peptide library. They let us as eukaryotic cells fine tune protein production and effect to meet the ever changing demands of the cellular environment. We are finding that microRNA probably represents the single largest quantity of RNA found in most cells. Comprehensive Written Questions (12 points) Choose 4 of the following 6 questions (3 pts each) Please write answers on the paper provided Useful Formulas: Keq= [products]/[reactants] R=1.987 cal/mol K T=298 K MM kinetics: v= Vmax [S]/(Km+[S]) LB Kinetics: 1/v= (Km/Vm)(1/[S]) + 1/Vmax ∆G= ∆G’o + 2.303 RT log [products]/[reactants] Remember!!!!! 2.302(log X) = ln X 59) Consider this reaction in a skeletal muscle cell: Phosphocreatine + ADP Creatine + ATP Phosphocreatine= 28 mM, Creatine = 8 mM, ADP=1 mM, ATP=8 mM, PO4=8mM A) What is the standard ∆G’o for this reaction? Show math, and final mathematical answer in CORRECT UNITS in correct formula. B) Show the formula with numbers set in proper units that would permit you to properly calculate the physiological delta G for the reaction under the conditions described above if you have access to a good calculator See Notes and Prior Delta G assignments 62) Draw and label the Michealis-Menton Plot for an enzyme-linked reaction and show the relationship of Vmax and Km. A) Normal Reaction (Km and Vm) B) Plot of reaction in the presence of a competitive inhibitor (Km ci and Vm ci) C) Plot of reaction in presence of a non-competitive inhibitor (Km nci and VM nci) D) Can an enzymes Vm ever in practice be reached? Yes or No Draw a plot like this on your answer sheet Rxn See Notes for three different curves Velo city [S] 63) What is the Fluid Mosaic Model of a cell membrane? What are the characteristics of membrane lipids and proteins that make this model work? (20-40 words with diagrams if this helps) This model states that the plasma membrane consists of a phopholipid bilayer with polar heads facing water (out) and the fatty acid tails facing each other in the hydrophobic center. Membrane proteins either span the bilayer (transmembrane) or associate with just the cytosolic or extracellular sides (integral or lipid-associated). Within each monolayer, phospholipids and proteins can float about or diffuse laterally across their respective phospholipid monolayers, however they are unable to change sides (flip-flop) because their polar ends or polar amino acids are unable to pass through the hydrophobic center of the bilayer. One limitation is that some proteins are securely anchored to intracellular structures. Ankyrin, spectrin and actin can all participate in this regard. This way some proteins are expressed on only one part of a cell’s plasma membrane. 64) With respect to glycolysis, many of the chemical reactions have a positive ∆G’o value. Describe three things that a cell could use to improve product formation? (20-40 words) Best: Remove product to pull substrates forward Another good one: Increase Substrate concentrations One fine trick: Link an unfavored reaction (+ΔG) with a favorable reaction (-ΔG) I nthe same compartment/close proximity (no membranes between reactions) Another fine trick: Use allosteric regulation where the product of a reaction or reaction pathway is used to change enzyme function when you need more or less product by binding non-covalently at an allosteric site Other options: Change temperature, Km values, make/destroy enzymes, change [P] or [S] concentrations, pH where reaction occurs, etc. 65) With respect to second messenger systems, why is intracellular amplification needed so that a hormone such as insulin can change cellular function? Name and describe a hormone that does not require amplification and how it functions in this regard. (20-40 words with diagrams if this helps) Some hormones are unable to enter the cytosol and otherwise unable to modify the intracellular functions of a cell. Other hormones are produced in such small amounts that they would be unable to create changes in cellular function. Fro these hormones amplification of the extracellular message is made possible by causing intracellular enzymes to change their function. For example insulin stimulates a tyrosine kinase on the intracellular side which then potentially phosphorylates thousands of proteins inside the cell, each target enzyme if a kinase or phosphatase can in turn modify the activity of thousands of other proteins. Testosterone and thyroxine are examples of hormones that do nto always cause or require amplification inside the cell. These hormones tend to be lipid soluble and can, though do not always, directly bind to DNA and alter the transcription of DNA into mRNA (enhancing or suppressing transcription and translation). Sometimes after entering the cytosol a nuclear binding protein will carry the hormoneprotein complex to the nucleus where it will exert its effects. 66) Consider microtubules and microfilaments: compare and contrast their structure (subunits) and function (describe two cellular functions that each is responsible for). (20-40 words with diagrams if this helps) Microtubules are the largest of the cytoskeletal systems and straight hollow cylinders Have an outer diameter of 25 nm Inner diameter of 15 nm Two general groups of microtubules in eukaryotic cells: Axonemal or Cytoplasmic microtubules Microtubules structurally are straight hollow cylinders composed of protofilaments Form a protofilament using simple monomers with Multiple (13 in total) protofilaments must interact to form cylindrical structure. The basic unit (monomer) of a protofilament is tubulin (α-tubulin or Βtubulin ) Unit Extra Credit (2 pts 4 X ½ pt): Draw/Name and amino acid that would do the following: a) Associate with the hydrophobic core of the lipid bilayer b) Could be phosphorylated by a kinase c) Would have a charged R-group that could attract calcium d) Would permit collagen to form tight bundles due to its small size See Notes Comprehensive Extra Credit (up to 4 pts): Create a diagram/flow chart that names the substrates, products, organelles and enzyme complexes that permit a single glucose molecule in the cytosol of a well oxygenated eukaryotic cell (cardiac cell) to be completely oxidized in a mitochondria to carbon dioxide, generate a proton gradient for ATP synthesis, as well as consume oxygen and ultimately produce of water. (chemical structures optional) See Notes