Advanced Cell Biology. Lecture 28
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1 Advanced Cell Biology. Lecture 28 Alexey Shipunov Minot State University April 8, 2013 Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
2 Outline Questions and answers Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
3 Outline Questions and answers Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
4 Questions and answers Previous final question: the answer How to knockout a gene? Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
5 Questions and answers Previous final question: the answer How to knockout a gene? Recombine with non-working (plasmid, homologous recombination etc.) Suppress Use RNA interference Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
6 Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
7 Selective barriers : Receive information Import and export molecules Have a role in moving and expanding cell Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
8 Cellular compartments Prokaryotes are one-membrane cells Eukaryotes have multiple closed and open one- and double-membrane compartments Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
9 Eukaryote cell compartments Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
10 Lipid bilayer Two layers of lipids Scattered proteins Carbohydrates attached to outer surface (cortex) Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
11 Amphipathic lipids Phospholipids Sterols Glycolipids Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
12 Amphipatic membrane lipids Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
13 Membrane lipids movie Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
14 Fluids Physically, membranes are lipid liquid suspension in water In artificial conditions, liposomes have structure similar to membranes Coacervate theory of life origin based on liposome existence Cholesterol can stiff membranes Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
15 Membrane fluidity movie Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
16 Liposomes Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
17 Why bilayers make liposomes Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
18 Asymmetry Different types of phospholipids located unevenly between two layers Enzyme flippase transfer lipids between layers Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
19 Saving membrane asymmetry Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
20 Membrane proteins: functional groups Transporters: Na + pump Anchors: integrines Receptors: PDGF (platelet-derived growth factor) receptor Enzymes: adenylyl cyclase (catalyze camp production) Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
21 Functional groups of membrane proteins Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
22 Membrane proteins: positional groups Transmembrane (integral, IMP) Monolayer-associated (IMP) Lipid-linked (IMP) Protein-attached (peripheral, PMP) Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
23 Positional groups of membrane proteins Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
24 Membrane proteins: secondary structure Mostly α-helices Sometimes also β-barrels (porins) Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
25 Membrane α-helix Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
26 Porin: β-barrel Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
27 Detergents Small lipid-like molecules Solubilize phospholipids, leaving proteins intact Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
28 Membrane disruption movie Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
29 Bacteriorhodopsin Small protein of 250 amino acids Proton pump activated by light Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
30 Bacteriorodopsin Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
31 Bacteriorodopsin movie Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
32 Cell cortex Cell cortex proteins like spectrins form a supportive network This network will stabilize a lipid liquid movements Some peptides may be restricted to particular domains of membrane Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
33 Spectrins Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
34 Plasma membrane domains Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
35 Membrane (cortex) carbohydrates Glycoproteins and peptidoglycans Form carbohydrate layer Protect, lubricate and used as recognition molecules (e.g., for lectins) Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
36 Cortex Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
37 Lectins Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
38 Some methods of membrane research FRAP (fluorescence recovery after bleaching) SPT (single-particle tracking) microscopy Solubilizing and reconstituting of membrane proteins Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
39 FRAP movie Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
40 Solubilizing and reconstituting Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
41 Final question (2 points) Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
42 Final question (2 points) What is the difference between receptor and transporter proteins? Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
43 Summary are selective, bilayer, fluid barriers Membrane lipids are amphipatic Membrane proteins do trans-membrane jobs Membrane (cortex) carbohydrates are protectives and informers Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
44 For Further Reading A. Shipunov. Advanced Cell Biology [Electronic resource] onwards. Mode of access: B. Alberts et al. Essential Cell Biology. 3rd edition. Garland Science, Chapter 11. Shipunov (MSU) Advanced Cell Biology. Lecture 28 April 8, / 41
Advanced Cell Biology. Lecture 28
Alexey Shipunov Minot State University March 30, 2012 Outline Questions and answers Outline Questions and answers Questions and answers Previous final question: the answer How to make a transgenic organism
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