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MODULE: Molecular Biology & Biochemistry MODULE NUMBER: BIO00004C JACS CODE: C700 STAGE/YEAR: 1 CREDITS: 20 ORGANISER: Dr Set Chong SUBJECT COMMITTEE: Biochemistry VERSION: 12 Nov 2015 TERMS TAUGHT: Au,Sp,Su 2016/17 PREREQUISITES: Students should have met the admissions standards required by the Department of Biology for enrolment on the course. RECOMMENDATIONS: AIMS This module deals with the structure and function of fundamental chemical molecules of a cell. It starts with an overview of the module and advanced applications that depend on the subjects covered in the module. It then covers the basic chemical building blocks of cells, from elements to macromolecules. The structure of nucleic acids will be introduced, and its importance to the mechanism of DNA replication. Then, the different levels of protein structure will be defined and protein-protein interactions, covalent modification and the nature of membrane proteins, described. The functionality of proteins as enzymes will be discussed in detail. Following an introduction to lipid and carbohydrate structures, the role of the various macromolecules in the context of membrane flow, cell shape, etc. will be discussed. Energy and metabolism is introduced by discussing the important concept of free energy and relating this to the central role of ATP and coupling of biochemical processes. The course then surveys carbohydrate and fat metabolism, photosynthesis and related metabolic processes in plants, and concludes with section on the integration of metabolism. LEARNING OUTCOMES: 1. To be able to describe the main chemical components of cells, their structural properties, how these relate to their functions, and how they are altered during cellular processes 2. To be able to describe and explain how covalent and non-covalent interactions bring about the assembly of cellular components and macromolecules 3. To be able to explain theoretical frameworks (such a Michaelis Menten kinetics, the laws of thermodynamics and the chemiosmotic theory) that allow us to understand function of biological molecules and cells 4. To be able to integrate knowledge about heterotrophic metabolism (of carbohydrates & lipids) and phototrophic metabolism and how they relate to energy metabolism via ATP 5. To be able to relate knowledge of biological molecules to health and disease and to their application in biotechnology 6. To be able to apply quantitative approaches to perform basic calculations related to acid-base chemistry, redox reactions, and to analyse and evaluate enzyme kinetics data gathered in practical classes SYNOPSIS OF TEACHING: Event Duration (Hrs) Topic Staff Room type Timing Lecture 1 1 Introduction. Highlights in Molecular Biology and Biochemistry. Peter McGlynn L Autumn term Lectures 2 and 3 1,1 Chemical composition of living things. Acids and bases. That macromolecules are made of building blocks. Covalent and noncovalent assemblages. Set Chong L “ Lectures 4 and 5 1,1 Structural features of DNA and RNA. Set Chong L “ Lectures 6,7,8,9,10 1,1,1,1,1 Proteins are the molecules that carry out active functions in the cell. Peter McGlynn Roles of amino acids, and levels of organisation in the structure of proteins (primary, secondary, tertiary, quaternary). Different types of non-covalent interactions. Protein folding (and disease). Co-factors / covalent modification. L “ Lectures 11,12 1,1 The enzyme in the cell, catalysis and specificity, active sites, stereospecificity, flexibility, conformational changes, induced-fit. Enzyme assays and enzyme kinetics. Classification of enzymes. Moir L “ Lecture 13 1 Serine proteases: control, mechanism, specificity and evolutionary relationships within an extensive enzyme family. Moir L “ Lecture 14 1 Allosteric enzymes and the control of enzyme activity. Cooperativity Moir and feedback inhibition, aspartate transcarbamylase as a case study. Survey of other control mechanisms. L “ Lecture 15 1 Protein/enzyme engineering; methods for designing new and modified enzymes, studies with subtilisin. Applications of enzymes in biotechology. Moir L “ Workshop 2hrs Solving problems related to material taught in Autumn term Split class to run (basic calculations, pH, etc.) See worksheet on the VLE. Each student does one 2-hr workshop. Moir, Peter McGlynn, Workshop Week 10 Practical /Lecture (1) 1 Set Chong L – where tbc Must be after L10 Practical 1 Duration 3 hours, Separation of biomolecules by column chromatography Set Chong Where tbc Must be after L10 & Practical L1 Lectures 16,17,18 1,1,1 Lipids, membrane bilayers. Carbohydrates: Monosacharides and polysaccharides. Cell walls and extracellular matrix. Roles of lipids and proteins in membrane function. Baumann L Spring Lecture 19 1 Introduction to energetics: free energy and the central role of ATP Maathuis as energy currency. Redox potentials and their relationship to Gibbs free energy. L “ Introduction to practicals[1] Lectures 20,21,22, 23,24 1,1,1,1,1 Metabolism. Overview and concepts of metabolism; how cells utilize Ungar different sugar nutrients, regulation of glycogen metabolism; detailed discussion of glycolysis, the TCA cycle and the role of fermentation; regulation of these pathways, how energy generation is balanced with the efficient turnover of available nutrients; how and why cells choose one pathway over another; generation of a redox currency for biosynthetic use – the pentose phosphate pathway; select examples of metabolic aspects of diseases. L “ Lectures 25, 26 1,1 ( i) The cell: control of glycolysis, the glucose/fatty acid cycle and fatty acid oxidation. (ii) The tissue: examples of the specialisation of carbohydrate and fat metabolism in various tissues. (iii) The organism - whole body co-ordination of metabolism by hormones: eg obesity and starvation L “ Lectures 27, 28 1,1 Conservation of free energy from carbohydrate catabolism as Maathuis NADH- quantitative assessment. The pathway of electrons from NADH to oxygen: technical approaches; functions of the major protein complexes; prosthetic groups in the electron-transporting complexes. Coupling electron transport to ATP synthesis and identification of those complexes involved in ATP synthesis. Measurement of P:O ratios. How ATP is synthesised: chemiosmotic coupling. Quantifying the energy in transmembrane solute gradients. The protonmotive force in coupled mitochondria; proton flows, stoichiometry and energetics. Proton flow and the phenomenology of redox-phosphorylation coupling. Ubiquity of the ATP synthase at “energy-coupling” membranes. Alternative uses or the PMF in bacteria: solute uptake and swimming. L “ Lectures 29, 30, 31 1,1,1 Photosynthesis. Transduction of light energy in chloroplasts. The Maathuis nature of light. The energy in a photon. Comparison of light energy with other forms of free energy. Pigments, absorption spectra and excited states. “Light-harvesting” and resonance energy transfer; antenna and reaction centre chlorophylls. Loss of energy in excited reaction centres and generation of redox reactions. Serial arrangement of photosystems I and II. Redox reactions and carbon fixation in chloroplasts. The components of the redox chain. The PSII reaction centre; cytochrome b6f complex; the PSI reaction centre. Cyclic phosphorylation. Carbon fixation, the Calvin cycle and pathways of starch and sucrose synthesis; photorespiration and C4 metabolism. L “ Practical (2-day) 1 x 4hr, 2 x 3hr, 1 x 1hr (PC to follow P3), 3 occurances Each student attends a 2 day practical (4x3 hr sessions) on the kinetics of the enzyme alkaline phosphate. Development of a spectroscopic assay; systematic investigation of how rates depend on enzyme and substrate concentration; determination of kinetic parameters Km and Vmax; studies of enzyme inhibition. This practical involves both experimental design, and some data handling. Moir Where tbc Spring, when tbc Workshop 2 2 hours Solving problems related to material taught in Spring term (lipids, carbohydrates, metabolism, energy, photosynthesis). Baumann, Maathuis, Ungar Where tbc Summer term, timing tbc Ungar Event Duration (Hrs) Topic Staff ASSESSMENT: Summative: Closed examination (1.5 hrs) in January assessment period (week 1, spring term), weighted 40% of module mark Closed examination (1.5 hrs) in summer assessment period (weeks 5-7 summer term), weighted 40% of module mark Open assessment, spring term, weighted 20% of module mark Re-assessment: Closed examination paper, August resit week DEMONSTRATING REQUIREMENTS: Autumn: 4 demonstrators Spring: A total of 4-6 demonstrators needed for all of the practicals, two demonstrators being on duty at any time. MAXIMUM NUMBERS: To capacity of lecture theatre/labs STUDENT WORKLOAD: students’ workload totalling 100 hours per 10 credit module Lectures: 32 hrs Workshops: 4 hrs Supported learning sessions (see workshops) Practicals: 15 hrs Tutorials Total Contact hours: 51 hrs Assessments (formative and summative): 2x 1.5 hrs Private study: 146 hrs Room type Timing