HST.410J/6.021J Lecture 3

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1 Harvard-MIT Division of Health Sciences and Technology HST.410J: Projects in Microscale Engineering for the Life Sciences, Spring 2007 Course Directors: Prof. Dennis Freeman, Prof. Martha Gray, and Prof. Alexander Aranyosi HST.410J/6.021J Lecture 3 Escherichia Coli (E. Coli) Culture Medium February 13, 2007 Image removed due to copyright restrictions. Photograph of an E. coli cell. NH 4 Cl MgSO 4 KH 2 PO 4 Na 2 HPO 4 Glucose Water 1.0 g 0.13 g 3.0 g 6.0 g 4.0 g 1.0 L Generic Eukaryotic Animal Cell Culture Medium Essential Amino Acids (a dozen) Vitamins (eight) Image removed due to copyright restrictions. Salts (Na +, K +, Ca 2+, Mg 2+, Cl, PO 3 4, HCO 3 ) Glucose Serum Air CO 2 Food, water Waste Respiratory system Digestive system Circulatory system Cells Inputs to Organism air water food carbohydrates fats proteins Inputs to Cells oxygen water ions building block molecules sugars lipids amino acids

2 Water transport in digestive system Salivary glands Trachea Liver Pharynx Esophagus Stomach Pancreas Daily traffic 800 g food L water ingested daily 1.5 L saliva 2 L gastric secretions 0.5 L bile 7 L digestive fluids 1.5 L pancreatic secretions 1.5 L intestinal secretions 15 pounds of water (10% of body weight) secreted and reabsorbed daily Small intestine Rectum Anus Large intestine Images removed due to copyright restrictions. Images of inside an intestinal tract and an endothelial cell.

3 Microvilli Junctional complex Apical (mucosal) surface Lateral surface Intercellular Basal (serosal) Transcellular Paracellular transport through surface transport transport gap junctions lipid bilayer membrane 7 nm cell Concentration at a point in space and time volume V amount of substance in V concentration c(x,t) = lim V V 0

4 Flux at a point in space and time window area A φ(x,t) amount of substance flowing flux φ(x,t) = lim through test window A in Δt A 0 AΔt Δt 0 Fick's First Law CH 2 OH CH 2 OH CH 2 OH C O C O C H H OH H H H H H C C C C C OH H OH H OH O OH H C OH OH H OH OH C C C C C C OH H OH H OH H H D-glucose D-mannose D-galactose

5 CH 2 OH CH 2 OH C O O C H H H H H H C C C C OH H H OH OH OH OH OH C C C C H OH OH H D-glucose L-glucose integral membrane solute protein lipid bilayer membrane 7 nm cell Hydrophobicity Residue position

6 Hydrophobic Hydrophilic Outside Inside NH2 COOH Figures from Weiss, T. F. Cellular Biophysics, Image removed due to copyright restrictions. Please see figure 5 in Mueckler, Mike, and Carol Makepeace. "Cysteine-scanning Mutagenesis and Substituted Cysteine Accessibility Analysis of Transmembrane Segment 4 of the Glut1 Glucose Transporter." J Biol Chem 280 (2005):

7 Pumps ADP + P ATP 2 K + 3 Na + Inside Membrane Outside integral membrane protein lipid bilayer membrane pore 7 nm cell

8 Y. Jiang, A. Lee, J. Chen, V. Ruta, M. Cadene, B. Chalt, and R. MacKinnon (2003), Nature 423: Images removed due to copyright restrictions.

9 Intracellular Dissolution Transport Transport Carrier- Pumps and diffusion through through mediated through water gated ion transport lipid bilayer channels channels Membrane Extracellular

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