Activity #1 Biochemical Systems PDF

Activity #1 Biochemical Systems Prepared by: Julie Rose de Guzman, RMT, MLS (ASCPi)cm  Saccharomyces cerevisiae  cells appear as ovoid or ellipsoidal structures, surrounded by a rather thick cell wall ...

Activity #1 Biochemical Systems Prepared by: Julie Rose de Guzman, RMT, MLS (ASCPi)cm  Saccharomyces cerevisiae  cells appear as ovoid or ellipsoidal structures, surrounded by a rather thick cell wall  MAJOR COMPONENTS AND ORGANELLES A. CELL ENVELOPE B. CYTOPLASM AND CYTOSKELETON C. NUCLEUS D. ORGANELLE COMPARTMENT  Saccharomyces cerevisiae  MAJOR COMPONENTS AND ORGANELLES A. CELL ENVELOPE A.1 CELL WALL - yeast cell aggregation; agglutination A.2 PLASMA MEMBRANE  The primary functions of the yeast plasma membrane are: i. Physical protection of the cell. ii. Control of osmotic stability. iii. Control of cell wall biosynthesis. iv. Anchor for cytoskeletal compounds. v. Selective permeability barrier controlling compounds that enter or that leave the cell. Of prime importance in active transport of solutes is the activity of the plasma membrane proton-pumping ATPase vi. Transport-related functions in endocytosis and exocytosis. vii. Location of the components of signal transduction pathways. viii. Sites of cell-cell recognition and cell-cell adhesion Periplasmic space -comprises mainly secreted proteins that are unable to permeate the cell wall, such as invertase & phosphatase  Saccharomyces cerevisiae  MAJOR COMPONENTS AND ORGANELLES B. CYTOPLASM AND CYTOSKELETON B.1. MICROTUBULES - are conserved cytoskeletal elements  B.2. ACTIN- direct polarized cell growth and to segregate organelles prior to cell division.  B.3. MOTOR PROTEINS- provide the energy necessary for motility. Myosin & Kinesins- are proteins that are able to bind to polarized cytoskeletal filaments and use the energy derived from repeated cycles of ATP hydrolysis to move along them. Dynein- largest motor protein active in the movement of the mitotic spindle that must move into the narrow neck between the mother cell and the bud in order to segregate duplicated chromosomes accurately  Saccharomyces cerevisiae  MAJOR COMPONENTS AND ORGANELLES C. NUCLEUS C.1. NUCLEAR PORE- form the natural channels for exchange of components between the nucleus and cytosol, whereby export and import pathways can be distinguished. C.2. NUCLEOLUS- locates the rRNA genes, and is the site for the synthesis and processing of rRNA. It is also involved in the assembly of the ribosomal subunits and in pre-mRNA processing C.3. CHROMOSOMES- genetic element  Saccharomyces cerevisiae  MAJOR COMPONENTS AND ORGANELLES D. ORGANELLE COMPARTMENT D.1. ER- key organelle for all processes controlling the stability, modification, and transport of proteins. D.2. GA- processing and sorting events include synthesis and processing of complex biomolecules. D.3 VACUOLES- involved in intracellular protein trafficking and nonspecific intracellular proteolysis; vacuole is a “drain.” ✓ storage compartments ✓ Osmoregulation ✓ homeostatic regulation of cytosolic ion conc. and pH.  Saccharomyces cerevisiae  MAJOR COMPONENTS AND ORGANELLES D. ORGANELLE COMPARTMENT  D.4. MITOCHONDRIA- respiratory-competent organelles; will adopt different morphologies, depending on the conditions.  D.5 PEROXISOMES- also called microbodies; endowed with several metabolic functions that are of outstanding importance for cell viability A. Characteristics of Cells Suspension A Suspension B … after adding stain Suspension A Suspension B Methylene Blue is a cationic stain Uses: 1. For determining cell damage 2. Identification of Microorganisms 3. Identifying Nucleic Acids 4. Identifying RNA Sequences 5. Calculating viable cells in yeast sample 6. Identifying distinctions between bacterial, viral and fungal diseases 7. As diagnostic agent Balloon test MECHANISM OF FERMENTATION - both the rate and the total amount of glucose consumption were many times greater under anaerobic than aerobic conditions. “Pasteur effect” -The ATP yield from glycolysis under anaerobic conditions (2 ATP per molecule of glucose) is much smaller than that from the complete oxidation of glucose to CO2 under aerobic conditions (30 or 32 ATP per glucose) CONCEPT OF OSMOSIS: The passage of solvent through a semipermeable membrane separating a dilute solution from a more concentrated solution OSMOTIC PRESSURE- must be applied to prevent the net flow of solvent through a semipermeable membrane from a lower concentration to a higher concentration B. Osmosis RAOULT’S LAW- states that a solvent’s partial vapour pressure in a solution (or mixture) is equal or identical to the vapour pressure of the pure solvent multiplied by its mole fraction in the solution; related to vapor pressure estimation  LAWS OF OSMOTIC PRESSURES  van’tHoff’s Theory of  New Osmotic Pressure Osmotic Pressure formula osmotic force equilibrium formula: atmospheric pressure ∙ osmotic effective area on the solvent side = (atmospheric pressure + osmotic pressure) ∙ osmotic effective area on the solution side (a substance in solution behaves exactly like a gas, and the osmotic pressure of a dilute solution is equal to the pressure which the solute osmotic pressure as π would exert if it were a gas at the atmospheric pressure as p same temperature occupying the molar concentration of solution as [C𝑖 ]. same volume as the solution). osmotic effective area on the solution 1 − 𝑘[𝐶𝑖 ]) temperature at melting point of solvent T/ T0 Glucose transporters (GLUT) C. Dialysis  Laboratory set-up: CONCEPT OF DIALYSIS process in which a semipermeable membrane allows the passage of solvent, dissolved ions and small molecules but blocks the passage of colloidal- sized particles and large molecules RATIONALE... - The helical structure of Amylose forms a charge transfer (CT) complex with iodine, wherein iodine is present inside the spiral or helical structure of the Amylose. - in the charge transfer complex of polyiodide ions and Amylose, electrons absorb light energy and get excited to a higher energy level. The complementary color to the light energy absorbed by the charge transfer complex is perceived as a blue-black color by the human eye. Application in living system... Le Chatelier's principle This principle focuses on three ways in which we can change the conditions of a chemical reaction at equilibrium: (1) changing the concentration of one of the components of the reaction (2) changing the pressure on the system (3) changing the temperature at which the reaction is run. D. Diffusion Area Application in living system... BIOCHEMICAL SYSTEM