Post on 21-Jan-2016
Lipids II; Membranes
Andy HowardIntroductory Biochemistry
4 March 2008
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What we’ll discuss Lipids Phospholipids Plasmalogens Glycosphingolipids
Isoprenoids Steroids Other lipids
Membranes Bilayers Fluid mosaic model
Physical properties
Lipid Rafts
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Glycerophospholipids Also called phosphoglycerides Primary lipid constituents of membranes in most organisms
Simplest: phosphatides (3’phosphoesters)
Of greater significance: compounds in which phosphate is esterified both to glycerol and to something else with an —OH group on it
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Categories of glycerophospholipids Generally categorized first by the polar “head” group; secondarily by fatty acyl chains
Usually C-1 fatty acid is saturated
C-2 fatty acid is unsaturated
Think about structural consequences!
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Varieties of head groups
Variation on other phosphoester position
Ethanolamine (R1-4 = H) (—O—(CH2)2—NH3
+) Serine (R4 = COO-)(—O—CH2-CH-(COO-)—NH3
+) Methyl, dimethylethanolamine(—O—(CH2)2—NHm
+(CH3)2-m) Choline (R4=H, R1-3=CH3) (—O—(CH2)2—N(CH3)3
+) Glucose, glycerol . . .
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iClicker quiz question 1
What is the most common fatty acid in soybean triglycerides? (a) Hexadecanoate (b) Octadecanoate (c) cis,cis-9,12-octadecadienoate (d) all cis-5,8,11,14-eicosatetraeneoate
(e) None of the above
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iClicker quiz, question 2
Which set of fatty acids would you expect to melt on your breakfast table? (a) fatty acids derived from soybeans
(b) fatty acids derived from olives (c) fatty acids derived from beef fat
(d) fatty acids derived from bacteria
(e) either (c) or (d)
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iClicker quiz question 3 Suppose we constructed an artificial lipid bilayer of dipalmitoyl phosphatidylcholine (DPPC) and another artificial lipid bilayer of dioleyl phosphatidylcholine (DOPC).Which bilayer would be thicker? (a) the DPPC bilayer (b) the DOPC bilayer (c) neither; they would have the same thickness
(d) DOPC and DPPC will not produce stable bilayers
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Plasmalogens Another major class besides phosphatidates
C1 linked via cis-vinyl ether linkage. n.b. The textbook figure 9.9 is correct; but it appears opposite the text related to sphingolipids, which is confusing.
Ordinary fatty acyl esterification at C2
Phosphatidylethanolamine at C3
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Roles of phospholipids
Most important is in membranes that surround and actively isolate cells and organelles
Other phospholipids are secreted and are found as extracellular surfactants (detergents) in places where they’re needed, e.g. the surface of the lung
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Sphingolipids Second-most abundant membrane lipids in eukaryotes
Absent in most bacteria Backbone is sphingosine:unbranched C18 alcohol
More hydrophobic than phospholipids
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Varieties of sphingolipids
Ceramides sphingosine at glycerol C3 Fatty acid linked via amideat glycerol C2
Sphingomyelins C2 and C3 as in ceramides C1 has phosphocholine
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Cerebrosides & gangliosides
Cerebrosides Ceramides with one saccharide unit attached by -glycosidic linkage at C1 of glycerol
Galactocerebrosides common in nervous tissue
Gangliosides Anionic derivs of cerebrosides (NeuNAc) Provide surface markers for cell recognition and cell-cell communication
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Isoprenoids
Huge percentage of non-fatty-acid-based lipids are built up from isoprene units
Biosynthesis in 5 or 15 carbon building blocks reflects this
Steroids, vitamins, terpenes Involved in membrane function, signaling, feedback mechanisms, structural roles
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Steroids Molecules built up from ~30-carbon four-ring isoprenoid starting structure
Generally highly hydrophobic (1-3 polar groups in a large hydrocarbon); but can be derivatized into emulsifying forms
Cholesterol is basis for many of the others, both conceptually and syntheticallyCholesterol:Yes, you need to memorize this structure!
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Other lipids Waxes
nonpolar esters of long-chain fatty acids and long-chain monohydroxylic alcohols, e.g H3C(CH2)nCOO(CH2)mCH3
Waterproof, high-melting-point lipids
Eicosanoids oxygenated derivatives of C20
polyunsaturated fatty acids Involved in signaling, response to stressors
Non-membrane isoprenoids:vitamins, hormones, terpenes
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Membranes Fundamental biological mechanism for separating cells and organelles from one another
Highly selective barriers Based on phospholipid or sphingolipid bilayers
Contain many protein molecules too(50-75% by mass)
Often contain substantial cholesterol too:cf. modeling studies by H.L. Scott
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Bilayers Self-assembling roughly planar structures
Bilayer lipids are fully extended
Aqueous above and below, apolar within
Solvent
Solvent
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Fluid Mosaic Model
Membrane is dynamic Protein and lipids diffuse laterally;proteins generally slower than lipids
Some components don’t move as much as the others
Flip-flops much slower than lateral diffusion
Membranes are asymmetric Newly synthesized components added to inner leaflet
Slow transitions to upper leaflet(helped by flippases)
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Fluid Mosaic Model depicted
Courtesy C.Weaver, Menlo School
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Physical properties of membranes Strongly influenced by % saturated fatty acids: lower saturation means more fluidity at low temperatures
Cholesterol percentage matters too:disrupts ordered packing and increases fluidity (mostly)
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Lipid Rafts
Cholesterol tends to associate with sphingolipids because of their long saturated chains
Typical membrane has blob-like regions rich in cholesterol & sphingolipids surrounded by regions that are primarily phospholipids
The mobility of the cholesterol-rich regions leads to the term lipid raft
Still somewhat controversial
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Membrane Proteins Many proteins associate with membranes
But they do it in several ways Integral membrane proteins:considerable portion of protein is embedded in membrane
Peripheral membrane proteins:polar attachments to integral membrane proteins or polar groups of lipids
Lipid-anchored proteins:protein is covalently attached via a lipid anchor
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Integral(Transmembrane) Proteins Span bilayer completely
May have 1 membrane-spanning segment or several
Often isolated with detergents 7-transmembrane helical proteinsare very typical (e.g. bacteriorhodopsin)
Beta-barrels with pore down the center: porins
Drawings courtesy U.Texas
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Other Membrane Proteins Peripheral membrane proteins
Associate with one face of membrane Easier to disrupt membrane interaction
Lipid-anchored membrane proteins Protein-lipid covalent bond Often involves amide or ester bond to phospholipid
Others: cys—S—isoprenoid (prenyl) chain
Glycosyl phosphatidylinositol with glycans
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Membrane Transport
What goes through and what doesn’t?
Nonpolar gases (CO2, O2) diffuse
Hydrophobic molecules and small uncharged molecules mostly pass freely
Charged molecules blocked
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Transmembrane Traffic:Types of Transport (Table 9.3)Type Protein Saturable Movement
EnergyCarrier w/substr. Rel.to
conc. Input?DiffusionNo No Down NoChannels Yes No Down No & poresPassive Yes Yes Down No transportActive Yes Yes Up Yes
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Cartoons of transport types
From accessexcellence.org
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Thermodynamics ofpassive and active transport• If you think of the transport as a chemical reaction Ain Aout or Aout Ain
• It makes sense that the free energy equation would look like this:
• Gtransport = RTln([Ain]/[Aout])
• More complex with charges;see eqns. 9.4 through 9.6.
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Example Suppose [Aout] = 145 mM, [Ain] = 10 mM,T = body temp = 310K
Gtransport = RT ln[Ain]/[Aout]= 8.325 J mol-1K-1 * 310 K * ln(10/145)= -6.9 kJ mol-1
So the energies involved are moderate compared to ATP hydrolysis
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Charged species Charged species give rise to a factor that looks at charge difference as well as chemical potential (~concentration) difference
Most cells export cations so the inside of the cell is usually negatively charged relative to the outside
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Quantitative treatment of charge differences Membrane potential (in volts J/coul):
= in - out
(there’s an extra in eqn. 9.4) Gibbs free energy associated with change in electrical potential isGe = zFwhere z is the charge being transported and F is Faraday’s constant, 96485 JV-
1mol-1 Faraday’s constant is a fancy name for 1.
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Faraday’s constant Relating energy per moleto energy per coulomb:
Energy per mole of charges,e.g. 1 J mol-1, is1 J / (6.022*1023 charges)
Energy per coulomb, e.g, 1 V = 1 J coul-
1, is1 J / (6.241*1018 charges)
1 V / (J mol-1) =(1/(6.241*1018)) / (1/(6.022*1023) = 96485
So F = 96485 J V-1mol-1
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Total free energy change Typically we have both a chemical potential difference and an electrical potential difference so
Gtransport = RTln([Ain]/[Aout]) + zF Sometimes these two effects are opposite in sign, but not always
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Pores and channels
Transmembrane proteins with centralpassage for small molecules,possibly charged, to pass through Bacterial: pore. Usually only weakly selective
Eukaryote: channel. Highly selective. Usually the Gtransport is negative so they don’t require external energy sources
Gated channels: Passage can be switched on Highly selective, e.g. v(K+) >> v(Na+)
Rod MacKinnon
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Protein-facilitated passive transport All involve negative Gtransport
Uniport: one solute across Symport: two solutes, same direction Antiport: two solutes, opposite directions
Proteins that facilitate this are like enzymes in that they speed up reactions that would take place slowly anyhow
These proteins can be inhibited, reversibly or irreversibly
Diagram courtesySaint-Boniface U.
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Kinetics of passive transport Michaelis-Menten saturation kinetics:v0 = Vmax[S]out/(Ktr + [S]out)
Vmax is velocity achieved with fully saturated transporter
Ktr is analogous to Michaelis constant:it’s the [S]out value for which half-maximal velocity is achieved.
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Primary active transport
Energy source is usually ATP or light Energy source directly contributes to overcoming concentration gradient Bacteriorhodopsin: light energy used to drive protons against concentration and charge gradient to enable ATP production
P-glycoprotein: ATP-driven active transport of many nasties out of the cell
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Secondary active transport Active transport of one solute is coupled to passive transport of another
Net energetics is (just barely) favorable
Generally involves antiport Bacterial lactose influx driven by proton efflux
Sodium gradient often used in animals
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Complex case: Na+/K+
pump Typically [Kin] = 140mM, [Kout] = 5mM,[Nain] = 10 mM, [Naout] = 145mM.
ATP-driven transporter:3 Na+ out for 2 K+ inper molecule of ATP hydrolyzed
3Na out: 3*6.9 kJmol-1,2K in: 2*8.6 kJmol-1
= 37.9 kJ mol-1 needed, ~ one ATP
Diagram courtesy
Steve Cook
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What’s this used for? Sodium gets pumped back in in symport with glucose, driving uphill glucose transport
That’s a separate passive transport protein called GluT1
Diagram courtesy
Steve Cook
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How do we transport big molecules? Proteins and other big molecules often internalized or secreted by endocytosis or exocytosis
Special types of lipid vesicles created for transport
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Receptor-mediated endocytosis Bind macromolecule to specific receptor in plasma membrane
Membrane invaginates, forming a vesicle surrounding the bound molecules (still on the outside)
Vesicle fuses with endosome and a lysozome Inside the lysozyome, the foreign material and the receptor get degraded
… or ligand or receptor or both get recycled
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Example: LDL-cholesterol
Diagram courtesyGwen Childs, U.Arkansas for Medical Sciences
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Exocytosis
Materials to be secreted are enclosed in vesicles by the Golgi apparatus
Vesicles fuse with plasma membrane
Contents released into extracellular space
Diagram courtesy LinkPublishing.com
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Transducing signals Plasma membranes contain receptors that allow the cell to respond to chemical stimuli that can’t cross the membrane
Bacteria can detect chemicals:if something useful comes along,a signal is passed from the receptor to the flagella, enabling the bacterium to swim toward the source
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Multicellular signaling
Hormones, neurotransmitters, growth factors all can travel to target cells and produce receptor signals
We’ll discuss this in detail after the midterm
Diagram courtesy Science Creative Quarterly, U. British Columbia