C2006/F2402 '11 Outline for Lecture #4-- updated 01/27/11 07:32 PM
2011 Deborah Mowshowitz, Department of Biological Sciences,
Columbia University, New York NY
Handouts: 4-A. Types of junctions between cells and/or cells & ECM
4-B. Epithelial Cells -- see Becker fig. 17-7 (17-17)
4-C -- Comparison of Types of Transport -- Flow Diagram (top) & Chart (bottom)
Remember: All class handouts are online and in boxes on 7th floor of Mudd, outside Dr. M's office. (Most are on the course web site, but a few are on Courseworks, as indicated.)
PP shown at the start of class are on Courseworks. Here are Links to Selected Pictures:
1. RBC & Spectrin web: See first picture in
Note: In this article, the term 'protofilament' is used for a short MF, not an IF.
2. Picture of an epithelial cell from Lodish . (Similar to handout and Becker figures.)
I. Extracellular Matrix -- wrap up of major components
See notes of last lecture.
II. Cell-Cell (& Cell-ECM) Connective Structures -- For Pictures see Sadava figs. 6.6 & 6.7 (5.6 & 5.7), & Becker Ch. 17 -- exact figures listed below. For comparisons, see table below and/or Becker table 17-1 (17-3). For diagrams, see handout 4-A.
A. Basic types of Cell-Cell Junctions -- for overall view, see Becker fig 17-7 (17-17) & table below. For the molecular details (FYI only) see fig. 17-3 (17-13). For diagrams, see handout 4-A &/or texts.
1. Gap junctions -- Becker fig. 17-11 (17-21); Sadava 6.7 (5.7) & 15.19 (15.16).
2. FYI: Plasmodesmata -- Sadava fig. 7-21 (15.20) or Becker 17-26 (17-25). The plant's equivalent of a gap junction.
3. Tight Junctions -- Becker fig. 17-9 (17-19)
4. Adhesive (anchoring) junctions See Becker fig. 17-8 (17-18) & 17-3 (17-13) for molecular details (for your interest only). For a nice EM picture (& diagram) see http://www.vivo.colostate.edu/hbooks/cmb/cells/pmemb/junctions_a.html
For diagram of adherens junction see picture from Alberts. Also has nice pictures of desmosomes, gap junctions, etc. Two main types:
a. Desmosomes = Spot Junctions (with IF)
b. Adherens junctions
= belt junctions (with MF)
B. Summary Table of Animal Cell-Cell Junctions
Name of Junction
Important Structural Features
Connexons; form gated channel or pore
Small gap between cells
Passage of small molecules and ions (signaling & nutrition)
Fusion of ridges of protein in membrane; form tight seal
|Claudins||No gap at ridge.||
Water Tight Seal between cells; divide membrane into regions
Adhesive or Anchoring
Intracellular Plaques with filaments. Classified
Larger gap between cells
C. Cell - ECM junctions. Connect cytoskeleton to ECM or solid support. Resemble half of an adhesive junction (adherens junction or desmosome). See handout 4-A (Also Becker Fig. 17-22 (17-11). For another diagram see http://celljunctions.med.nyu.edu/images/figure1.gif.
1. Similarities to cell -cell adhesive junctions
a. Transmembrane protein connected (indirectly) to IF or MF on inside of cell.
b. Linker proteins involved -- linker proteins connect IF or MF to transmembrane protein.
2. Differences from cell-cell adhesive junctions
a. Transmembrane protein is an integrin, not a cadherin (See Becker fig. 17-21 (17-10) for a diagram of structure of integrin.) Integrins are a family of proteins with roles in signaling, movement, etc. as well as adhesion. (FYI: See Sadava, fig. 6.8, 9th ed, or Life of the Cell animation for signaling role of integrins.)
b. Transmembrane protein connects to ECM. (Not to extracellular domain of protein from another cell. )
3. Two Types.
Structure = "half desmosome";
Connect to IF on inside of cell.
Connect to basal lamina on outside.
Have plaque (thickening visible in EM) formed by linker proteins connecting to IF on inside of cell.
Usually found in epithelial cells.
b. Focal Adhesions
Structure = "half adherens junction"
Connect to MF on inside of cell.
Connect to ECM (can be on a solid support) on outside.
Connected to cytoskeleton by linker proteins, but no visible plaque.
Usually found in migrating cells.
At this point, it is a good idea to make a chart for yourself that classifies (or compares and contrasts) all the types of junctions, whether cell-cell or cell-ECM. Also try problems 1-12 & 1-14.
III. Types of Specialized Cells & an Example What does a real cell look like? Where are the junctions, etc.?
A. Cells & Tissues
1. Specialization. All cells in multicellular organism are specialized; there is no "typical cell."
2. Types. About 200 different cell types per human.
3. Tissue = Group of cells with similar structure & function that work as a unit.
Note: The word "tissue" is also used in a nonspecific way to mean a group of cells derived from an organ or system as in "kidney tissue." A kidney is an organ made up of many different tissue types.
4. 4 Major cell/tissue types -- muscle, nerve, connective, epithelial.
Note: Some people count blood as a separate category, so they get a total of 5 major cell types instead of 4.
B. The Four major Tissue Types (See Sadava fig. 40.7)
1. Muscle -- three kinds (skeletal, smooth & cardiac muscle); all specialized for contraction. (Sadava 40.4)
2. Nervous -- individual cell is called a neuron. Specialized for conduction of messages. (Sadava 40.6)
3. Connective -- cells dispersed in a matrix. (Sadava 40.5.) Extracellular matrix can be solid (as in bone), liquid (as in blood) or semi-solid (gel like) as in cartilage, adipose tissue. (Note fat in adipose tissue is stored inside the adipose cells in vesicles, not between the cells in the matrix.) See Sadava 40.4 in 6th ed. Connective tissue surrounds other tissues and provides support, protection and transport of materials to and from tissues (in blood).
4. Epithelial -- for an example see handout 4-B. For a similar diagram, see Becker fig. 17-7 (17-17). For a picture, see Sadava 40.3 (41. 3). For more pictures & diagrams, go to Google images or to the Pubmed bookshelf and search for 'epithelial cell junctions.' The Lodish text has a good picture that matches the handout.
a. Cells tightly joined -- use junctions described above to connect cells to each other and/or to ECM.
b. Make up linings of external and internal surfaces
c. Usually sheets. Can have one or more layers
d. Cells are Polarized -- two sides of cell layer are different. Contain different proteins and/or lipids in different domains (areas) of membrane.
(1). Terminology: Side of cell membrane facing lumen = apical surface; side facing body = basolateral (BL) surface.
(2). What keeps the two domains separate? Tight junctions.
e. Often rest on noncellular support material = basal lamina = part of ECM secreted by cells (on BL side)
f. Can form glands = epithelial tissues modified for secretion. (More details on formation, structure & function of glands later in course.)
g. Usual functions: selective absorption (selective transport across sheet), protection, secretion (from glands). See below for more on transport.
h. An example: epithelial layer surrounding the gut. See handout 4B or Becker Fig. 17-7 (17-17)
Now try problem 1-13 & 1-19. By now you should be able to do all the problems in problem set #1.
This leads to the next topic: How does the intestinal epithelium function in transport? How are substances are transported across membranes?
IV. Types of Transport Across Membranes (of small molecules/ions). For an overall summary, see table on handout 4-C. For reference, types of transport are numbered 1-5 on handouts & below. Also see Becker, fig. 8-2 or Sadava table 6.1 (5.1).
A. Classification of Transport -- three basic criteria for classifying transport
1. Role of transport protein (if any)
2. How energy is supplied
3. Direction things move (if more than one substance is moving)
B. Role of Protein -- Basic Types of transport classified by role of transport protein (if any). See handout 4-C.
1. No protein involved -- Simple Diffusion (case 1). Effective only for hydrophobic molecules (such as steroid hormones), gases, and very small molecules that can diffuse across lipid bilayers. See Becker table 8-1 & figure 8-5.
2. If Protein involved -- transport protein can be a channel, permease (carrier or exchanger) or pump. Cases 2-5.
a. channel (case 2) -- protein forms a pore allowing passage of hydrophilic materials across the lipid bilayer. Most, but not all channels, are ion channels.
b. transporter -- permease, carrier or pump -- protein binds to substance(s) on one side of bilayer, protein changes conformation and releases substance on other side of bilayer. Cases 3-5.
C. How energy is supplied to move X
1. Active vs. passive -- whether substances flow down their gradients (passive transport -- cases 1-3) or are pushed up their gradients by using energy (active transport -- cases 4 & 5). See Becker table 8-2 .
a. Passive transport -- substance moves down its concentration gradient. This can be by simple diffusion, passage through a channel, or with the help of a carrier protein. Cases 1- 3.
Membrane [X] high, → [X] low ΔG <0 (high concentration) (low concentration)
Note on terminology: Passive transport that requires a carrier protein (case 3) is usually called facilitated diffusion or carrier mediated transport. The term 'facilitated diffusion' is sometimes also used to refer to transport through a channel (case 2). The mechanisms in the two cases (2 vs 3) are distinctly different, so it is important to be careful about the distinction between carrier-mediated and channel-mediated transport.
b. Active transport -- substance moves up its concentration gradient with the help of "pump" or co-transporter protein and expenditure of energy (see below).
c. Transport can be in or out. Transport can be from the inside of the cell (or vesicle) to the outside or vice versa. In most of the examples discussed below, we are looking at transport into a cell (or vesicle), but transport out of cells is just as important.
2. Primary vs Secondary Active Transport -- See Becker fig. 8-9 or Sadava fig. 6.17 (5.15) for comparison of the 2 kinds of active transport.
a. Primary (or direct) active transport (Case 4) -- energy for transport is supplied by hydrolysis of ATP. In other words, the following two reactions (i) & (ii) are coupled:
Membrane (i). [X] low, out* → [X] high, in ΔG >0 (low concentration outside) (high concentration inside) (ii). ATP + H2O → ADP + Pi. ΔG <<0 ------------------------------------------------------------- Net: X out + ATP + H2O → X in + ADP + Pi ΔG <0 (X moves up its gradient at expense of ATP)
*Note: We are arbitrarily calling the two sides of the membrane 'in' and 'out.' What matters is that X is being moved from an area of low concentration to an area of higher concentration. (What happens in passive transport?)
Examples: pumps that move Ca++ into the ER or H+ into vesicles such as lysosomes. (The ER stores Ca++. Lysosomal enzymes work best at acid pH, unlike most other enzymes that work best at neutral pH.)
b. Secondary (or indirect) active transport (Case 5) -- energy for transport is supplied by some 2nd substance running down ITS gradient (reaction (ii) below). The following two reactions (i) & (iii) are coupled:
Membrane (i). [X] low, out → [X] high, in ΔG >0 (low concentration outside (high concentration inside) (iii). [Y] high
on one side of membrane
→ [Y] low
on other side of membrane
ΔG <<0 ------------------------------------------------------------- Net: X out + [Y] high → X in + [Y] low ΔG <0 (X moves up its gradient while Y flows down its gradient)
(1) Where is the high concentration of Y? Which way does Y move? See table below.
In some cases, Y moves in the same direction as X (symport).
In other case it moves in the opposite direction (antiport).
(2) An example: Glucose/Na+ co-transport -- glucose is pushed up its gradient by energy derived from Na+ going down its gradient. X = glucose; Y = Na+.
(3) Is ATP Involved? ATP is usually used to establish the gradient of [Y], but ATP is not directly involved here. For example, consider Glucose/Na+ co-transport.
The Na+/K+ pump (details next time) is used to establish a Na+ gradient. (This is primary active transport, and uses ATP.)
Once a Na+ gradient exists, the Na+ running down its gradient provides the energy to move glucose. (This is secondary active transport, and does not require ATP.)
D. Direction things move -- See Becker fig. 8-7 or Sadava fig. 6.15 (5.13)
|Type of Transport||
|One substance moves.||
Carrier mediated transport of Glucose; Ca++ transport into ER
|Two or more substances move in same direction.||
|Two or more substances move in opposite directions.||
Na+/K+ pump (active
-- driven by hydrolysis of ATP);
* An exchanger can be considered passive transport (carrier mediated), since the concentrations of the substances themselves drive the reaction . Alternatively, it can be considered secondary active transport, because movement of one of the substances down its gradient can drive transport of the other substance up its gradient.
E. Summary Table -- See Handout 4-C. For animations of transport done by Steve Berg at Winona State University go to Facilitated Diffusion Primary Active Transport. Berg's web site has many nice pictures & animations of cell and molecular processes.
Try problem 2-2.
V. How transport is measured (Probably Next Time)
A. Need a suitable experimental set up. A common method: using RBC ghosts.
B. How is it done?
Put ghosts in solution with some concentration of X on the outside and none on the inside.
You measure the amount of X that was transported inside the ghosts as a function of time. (At each time, you take a sample, centrifuge out the ghosts, and measure the amount of X in the ghosts.)
You repeat with different starting concentrations of [X]out
C. What do you learn from doing this?
You can tell active transport from passive.
You can infer the role of the transport protein -- you can tell if it is a channel or a carrier/pump.
How? Details in lecture 5!
Next Time: Important features of each type of transport, and an example of how the various types of transport are used. How glucose gets from lumen of intestine → muscle and adipose cells. Then, how do big molecules get into cells?