PHYSIOLOGY

Lecture: #1 Membrane Function & Structure, Basic Ion Principles Date: Wednesday, August 28, 2013 Time: 1PM Lecturer: Roman Shirokov, PhD Assigned reading: Chapter 1 ADMINISTRATIVE TOPICS

Lecture #1, 8/28

CLASS ANNOUNCEMENTS. At 1PM on Thursday, Dr. Stout will talk about the PA sessions. Also, we are working on getting the course listserv to work. You should have gotten 2 emails today from Dr. Stout via Pauline Castro. The email contains answers to questions you asked yesterday. I will take a poll tomorrow in class to see how many people have this working. CONTACT: Roman Shirokov, PHD OFFICE: I617 MSB EMAIL: roman.shirokov@umdnj.edu PHONE: 973-972-8869 (office), 973-972-0424 (lab) OPENING REMARKS FROM SHIROKOV. Everyone says that physiology is a hard course, but I will try to dispel that and make it easier to understand. Physiology is thought to be science of how organs function, however that is not correct. I present the molecular side of this science, from the physics of biology. Its inevitable that you have to start physiology from a physical/chemical route. Physiology is not like a pile of bricks, rather a system of letters that are interconnected. Every page of your lecture manual is loaded with ideas that will be built upon later in the course. You have to study every day and understand every word. OFFICE HOURS: None just contact him. There are no stupid questions, only stupid answers. =]

MEMBRANE PHYSIOLOGY This section about membranes and ions is the core for physiology- it will make the rest of this course much easier if you understand this part. (Slide. Membrane distribution in eukaryotic cells) Note that the plasma membrane makes up only a small percentage of the cell, for example 5% in pancreatic exocrine cell. Nevertheless, they cover a large surface area. IMPORTANCE OF MEMBRANES. To compartmentalize and maintain homeostasis (a fancy word for status quo). Membranes do not just compartmentalize proteins and organelles, they also

compartmentalize ions. The ion concentrations inside and outside the cell are very different and controlled. (Slide. Solute and electric charge imbalance) Outside the cell is sodium and chlorine ion rich. Inside is potassium rich. There are no free calcium ions inside the cell. Note: this table of ion concentrations is very important. It will come up several times. MAIN FUNCTIONS OF MEMBRANE: Compartmentalize (separate and control space) and transport TRANSPORT WHAT? TRANSPORT AREAS 1. Matter, such as sugar and water, 2. Information, such as signaling with ions (1) Signal transduction- across membrane (2) Propagation- along membrane

STRUCTURE OF THE MEMBRANE Glyco=sugar Lipid proper = membrane proper THE CELL MEMBRANE. It is a phospholipid bilayer that contains lipids and proteins. The cell coat is called the glycocalyx, formed of carbohydrates. The cell cortex contains actin proteins that are part of the cytoskeleton. LIPIDS IN MEMBRANE. Phospholipids (30-40%), the rest is: cholesterol, glycolipids, sphingolipids We are looking mainly at phospholipids that are glycerol based. If you cook up triacylglycerol in NaOH, you will make soap. UNSATURATED FAT. Fats that have at least one double bond in the hydrocarbon chain. These have lower melting points than saturated fats, because the double bond creates a kink in the chain. This kink causes more fluidity between other chains. AMPHIPHILICITY. Phospholipids are both hydrophobic (at nonpolar tails) and hydrophilic (charged head caused by phosphate group and hydrophilic group). CHOLESTEROL. Another important building block of membrane. Flat, rigid, slightly charged. GLYCOLIPIDS. Like phospholipids, they have hydrocarbon tails and a polar region. But no phosphate. (Slide. Intermolecular forces in solution) Note the +/- charges, indicating dipoles on the water molecules. The two long ovals are hydrophobic particles. Disturbing the hydrogen bonds between water molecules requires input of energy. Therefore, triglycerides like fat and oil stick together rather than mix with water- to avoid disrupting the network of water molecules. NOT A SOLUTION. Milk is a homogenized mixture of fat suspended in water.

MICELLES. Amphipathic phospholipids and glycolipids form micelles (spheres) in water to shield the hydrophobic tails from water. This is the same concept for cellular membranes- phospholipids form a bilayer that exposes the hydrophilic heads and keeps hydrophobic tails in the middle, like a sandwich. (3D molecular model of phospholipid.) (3D molecular model of cholesterol.) (3D molecular model of micelle.) (3D molecular model of phospholipid bilayer.) Channel protein is lodged in middle of hydrophobic tails, allowing a way for water to pass through the membrane. PROTEINS IN THE MEMBRANE (1) Peripheral proteins (2) Integral proteins-Transmembrane domain (single pass), Multiple TMD (multipass) You can release peripheral membranes from a cell membrane by changing the pH of the solution its in. This disrupts the bonding between peripheral protein and integral proteins contained in cell membrane. To release the integral proteins, you need detergent. MAJOR FUNCTIONAL GROUPS OF MEMBRANE PROTEINS. Transporter- transports ions and molecules in and out of cell Linker- attach to, stabilize, move membrane Marker- makes cell recognizable or attachable to other cells Receptor- send signal through signal transduction pathway in cell Enzyme- converts products in cell, happen to be located on membrane

GLYCOCALYX Functions to: 1. Protect cell from damage 2. Lubricate when it contacts other cells (Slide. Picture of microvilli) 3. Cell/cell recognition

COMBINATORIAL POTENTIAL. Since glycocalyx is made with sugars, there are many diverse combinations of sugars that can form unique tags for cell/cell recognition. EX. Red Blood cells

CYTOSKELETAL PROTEINS Functions to: 1. Give cell more rigid structure 2. Intracellular transport

*Note, no need to memorize Spectrin-Ankyrin cortex network slide.

EX. Red blood cells undergo compression when moving through capillaries. The Spectrin-Ankyrin cortex keeps the integrity of red blood cells.

SECOND HALF OF TODAYS LECTURE: IONS Key point: ions are separated by plasma membrane, resulting in different concentrations on inside and outside of cells MIXTURE. More than one substance SOLUTION. Mixture that is homogenous. Solution is a mix of molecules, components that have unique chemical properties. SOLVENT. Most abundant component in mixture SOLUTE. Substance dissolved in solvent. UNITS OF CONCENTRATION Volume solute/ volume solution x 100% = volume percentage or %v/v. However, we dont know how many molecules are in solution when reporting in %v/v. The information we need is molecular weight. MOLARITY. M = molar amount of solute / volume of solution (L) = mol/L AVOGADROS NUMBER. 6 x 10^23. Defined as how many molecules are in 12g of diamonds (carbon). 1 mol is 6 x 10^23 particles.

EXAMPLE WITH WATER MW (molecular weight) 2 x 1(H) + 16(O) = 18g/mole 1000 g/L 18g/mol Density= 1kg water/L water

= 55mol/L = 55M

All other molecules in the body are at least 100x less concentrated than water. For example, even though Na is relatively concentrated extracellularly, it is only 145mM = 0.145 M. 60% body weight is water 40% is intracellular fluid

20% extra cellular fluid

(Slide. Mass per volume concentration of protein) By weight, proteins make up a lot of mass per volume. It is 22% of cells!

DIFFUSION DIFFUSION. Diffusion is simplest transport mechanism. Particles go from regions of high concentration to low concentration. It is caused by the random motion of molecules. This is referred to as moving down the concentration gradient. FLUX. Flux is directly proportional to concentration difference of molecules. This is Ficks Law. Flux = -D (diffusion coefficient) x difference in concentration / difference in membrane thickness D is approximately the inverse of radius or molecular weight. Larger particles move more slowly. Flux is also affected by membrane permeability, the proportionality constant between flux and gradient PERMEABILITY. How easily substance passes through membrane Permeability is very low for large polar uncharged molecules, ions, and polar charged molecules (amino acids, nucleotides, phosphate). Low for water and small uncharged polar molecules. High for small gases like oxygen and carbon dioxide, because these are nonpolar. PARTIION COEFFICIENT (Beta) tells you how soluble something is in lipids, aka. How lipophilic. Flux = -Permeability x change in concentration Xylocaine and novocaine are two local anesthetics with high partition coefficient. As a result, they diffuse through cell membranes easily. OSMOSIS Dr. Shirokov is starting some of tomorrows lecture now. OSMOSIS. Caused by diffusion of water down its concentration gradient. Osmosis occurs across biological membranes in the presence of special water channels, called aquaporins. It happens when membranes have selective permeability, when the solute cannot cross the membrane, but water can. (Slide. Osmosis is caused by the diffusion of water) Note that the water-permeable membrane does not allow sugar to diffuse, since the molecule is too big. This is an example of selective permeability. EQUILIBRIUM is reached when there is no net movement of water. The flux of water in one direction equals the flux of water in the opposite direction. This does not mean there is no movement, just that there is no net movement.

HYDROSTATIC PRESSURE. When the force of hydrostatic pressure (caused by build of a water column on the right side) is equal to the force of water diffusion, there will be no more net movement. OSMOTIC PRESSURE. The osmotic pressure of solutes is the amount of pressure that stops net flux of water, in other words stops osmosis. Measured under constant volume condition. It is proportional to the concentration difference of solutes. P osmotic ~ N osm/ V osm

VANT HOFF LAW P osm = n RT / V P osm = RTC n: moles of solute R: gas constant T: absolute temperature (K) C: osmolarity of solute