BIOC*4580 Membrane Biochemistry Winter 2013

Lecture 2 Structures of Membrane Lipids Membrane Lipids (I)

Dr. George Harauz Molecular & Cellular Biology University of Guelph gharauz@uoguelph.ca

Review
Permeability, selfassembly, diversity of composition, asymmetry, , Unifying model!

Themes for Todays Lecture

Membrane lipids are highly diverse but share the common property of being AMPHIPATHIC They can be classified in various ways depending on headgroup and fatty acid composition, for example, and we shall describe major structural lipids Lipidomics is the process of determining the precise lipid composition of a cell and involves step-wise procedures of chemical fractionation and identification by mass spectrometry Synthetic lipids are often used by biochemists and biophysicists because of their well-defined properties

General Categories of Biological Lipids Diversity!

For today, just storage & structural lipids.

0. Fatty Acids
Nomenclature

Biological macromolecules are made of fundamental building blocks

For lipids, these are FATTY ACIDS.

Naming conventions of fatty acids 2x

Nomenclature of some naturally occurring fatty acids in animals

Number of carbons 12 14 16 16 18 18 18 18 20 Number of double bonds 0 0 0 1 0 1 2 3 0 Common name Laurate Myristate Palmitate Palmitoleate Stearate Oleate Linoleate Linolenate Arachidate Formula CH3(CH2)10COOCH3(CH2)12COOCH3(CH2)14COOCH3(CH2)5CH=CH(CH2)7COOCH3(CH2)16COOCH3(CH2)7CH=CH(CH2)7COOCH3(CH2)4(CH=CHCH2)2(CH)6COOCH3CH2(CH=CHCH2)3(CH)6COOCH3(CH2)18COO-

cf., Lehninger Table 10-1.

Measuring Hydrophobicity of Fatty Acids

Partitioning experiment using 2 solvents, e.g., heptane & water Measure partition coefficient:
Kp= Keq = [solute]H2O / [solute]heptane

Calculate free energy change:

Gtr = -RT ln(Keq)

(Structural) membrane lipids

Different classes of lipids:
Phospholipids Glycolipids Ether lipids Sterols

Diversity is the name of the game Asymmetry of lipid (& protein & carbohydrate) composition is, too

Categories of membrane lipids

PINK backbone (glycerol or sphingosine) YELLOW long-chain alkyls BLUE polar head group DIVERSITY from combinations of FA tails & polar heads

1. Phospholipids
1st Glycerophospholipids 2nd Sphingolipids

Phospholipids Glycerophospholipids 1st

Phospholipids have either a glycerol or sphingosine backbone first glycerophospholipids.

Glycerophospholipids (= Phosphoglycerides)

Not -5

Common Glycerophospholipids

Common Glycerophospholipids

PI(4,5)P2 has a theoretical net charge of -5 (and not -4 as in Lehninger Figure 10-9) but in reality each phosphate ester is only -1.5 (partially-ionized OH groups)

Summary Phospholipids with Glycerol Backbone

Phosphoglycerides or glycerophospholipids Acyl chains glycerol phosphate alcohol
(similar for all) (variable)

Common Phospholipids with Glycerol Backbone (to know!)

Full name
Phosphatidic acid Phosphatidylethanolamine Phosphatidylcholine Phosphatidylserine Phosphatidylinositol Phosphatidylglycerol Diphosphatidylglycerol (cardiolipin)

Abbreviation PA PE PC PS PI PG DPG or CL

Backbone charge -1 -1 -1 -1 -1 -1 -2

Headgroup charge 0 +1 +1 +1/-1 0 0 0

Net charge Anionic Zwitterionic Zwitterionic Anionic Anionic Anionic Anionic

Phospholipids Sphingolipids 2nd

Phospholipids have either a glycerol or sphingosine backbone second sphingolipids.

Phospholipids of the sphingolipid type

PC versus SPM

PC & SPM look pretty similar as spacefilling models.

Summary Phospholipids with Sphingosine Backbone

found in animal plasma membranes sphingosine + N-acyl fatty acid = ceramide Sphingomyelin SM Features: Phosphocholine headgroup (like PC) Sphingosine + fatty acid in amide link Zwitterionic (+ & -)

2. Glycolipids
1st Glycerol backbone (plant chloroplasts) 2nd Glycosphingolipids

Glycolipids

Glycolipids also have either a glycerol or sphingosine backbone, but lack the characteristic phosphate of phospholipids.

Plants:
Neutral Glycolipids (=Galactolipids) of chloroplast thylakoids

Sulpholipids (-vely charged)

(Phospholipids of the sphingolipid type)

Glycolipids of the sphingolipid type

(Table 7-1 has symbols for monosaccharides)

Ganglioside GM2 (mono series)

Wikipedia, alas, & Lehninger p 354

Summary Glycolipids with Glycerol Backbone

A) glycerol backbone: not commonly found in animal cells which have predominantly glycerophospholipids Plant chloroplast (thylakoid) membranes:
monogalactosyldiacylglyceride digalactosyldiacylglyceride sulfolipids with negatively-charged headgroup

others found in bacteria, etc.

Summary Glycolipids with Sphingosine Backbone

B) compare phospholipids with sphingosine backbone: - found in animal plasma membranes - sphingosine + N-acyl fatty acid = ceramide - sphingomyelin = ceramide + phosphocholine

versus: glycolipids with sphingosine backbone: ceramide + oligosaccharide = glycosylceramide (generic) neutral glycosphingolipids
uncharged sugars, e.g., galactosylceramide, lactosylceramide

acidic glycosphingolipids
Gangliosides Contain 9C acidic sugar: N-acetylneuraminic acid (sialic acid)

3. Ether versus Ester Lipids

Mainly archaeal, also animal

(Archaeal) Ether Lipids

Hydrocarbons attached to glycerol via ETHER linkage Stable at low pH & high T Archaea but also some animal tissues

Ester versus Ether Linkages

Ester alcohol + acid

Ether dehydration of alcohols

Wikipedia

Ether lipids in Archaea

Tetra-ether lipids span entire membrane Diphytanyl chains (40C, 8 x 5-C branched isoprene units) Linked to 2 polar headgroups, one in each leaflet

Ether lipids plasmalogens (enriched in heart muscle)

Ether lipids PAF (platelet activating factor a molecular signal)

Summary Ether Lipids

some animal tissues and unicellular organisms contain lipids in which hydrocarbon chains are attached to glycerol via an ether link, as either a C1-C2 alkene, or an alkane highly stable linkage at low pH and high temperatures; resistant to phospholipase degradation (see later) e.g., plasmalogens; enriched in heart muscle e.g., platelet-activating factor; released from leukocytes e.g., archaebacteria; contain tetraether lipids - long diphytanyl chains (40C, eight 5-C branched isoprene units) span the entire membrane and are linked to two polar headgroups, one in each leaflet

4. Sterols
Cholesterol (also stigmasterol & ergosterol), & derivatives thereof

Introducing the Sterols

Cholesterol modulates membrane fluidity in animal cell membranes but not in bacteria. Plants & fungi have alternative sterol derivatives.

Cholesterol rigid, planar, amphipathic Plants have, e.g., stigmasterol Fungi have ergosterol Bacteria have none

Figure 2.9 Close packing of cholesterol with lipids shown by MD simulations

Cholesterol cannot form bilayers alone but intercalates into membranes where it modulates fluidity.

Luckey Membrane Structural Biology

Steroids from Cholesterol

Structural Aspects of Membrane Lipids

Amphipathicity, nomenclature of FA chains (again), lipid variability & asymmetry

Common structural features of membrane lipids

Saturated versus unsaturated FA chains straight versus kinked

Nomenclature Fatty Acyl Chains & Phospholipids

# of C : # of = (double bonds) 14:0 16:0 18:0 18:1 18:2 Name myristate palmitate stearate oleate linoleate Property Saturated Unsaturated

> 100 individual phospholipid species in typical membrane Some synthetic single acyl chain type phospholipids: DMPC (dimyristoyl-PC) DPPE (dipalmitoyl-PE) DOPS (dioleoyl-PS) Mixed acyl chains found in natural membranes: SOPC (1-stearoyl-2-oleoyl)

Synthetic (DMPC, DPPE, DOPS) & natural (SOPC) phospholipids

DMPC DPPE DOPS SOPC
www.avantilipids.com

There are significant differences in lipid composition amongst membranes from different species & tissues
Lipid PA PC PE PG PI PS Cardiolipin Sphingomyelin Glycolipids Cholesterol Human Erythrocyte 1.5 19 18 0 1 8.5 0 17.5 10 25 Human Myelin Beef Heart Mitochondria 0.5 10 20 0 1 8.5 0 8.5 26 26 0 39 27 0 7 0.5 22.5 0 0 3 E. Coli 0 0 65 18 0 0 12 0 0 0

Weight percentages of total lipid

The different constituent membranes of the rat hepatocyte contain different lipids

n.b., high variability in Cholesterol!

Within each membrane, there is also asymmetry between leaflets

Summary of Lipid Diversity and Asymmetry

inner and outer leaflets have different lipid composition true for plasma membrane and probably all organelle membranes e.g. erythrocyte membrane
total phospholipid evenly distributed PC and SM (choline phospholipids) predominantly in outer leaflet PE and PS (amino phospholipids) inner leaflet

presence of PS in outer leaflet indicates that a cell is dead or dying, and marks it for destruction glycolipids are found only in the outer leaflet

STRUCTURES OF MEMBRANE LIPIDS YOU SHOULD BE ABLE TO DRAW

Fatty acids: know most of those listed in Table 12.1 (particularly of sizes 12-18 carbons, compare with Table 10.1 for full names) in the text Phospholipids: PA, PC, PE, PS, PG, PI, PIP2, cardiolipin Also know headgroup charges, etc. Sphingolipids: ceramide, SM Cholesterol: headgroup + ring structure + acyl chain Glycolipids (general idea): monogalactosyl- and digalactosyldiglyceride galactosylceramide, glucosylceramide General idea of the others (oligosaccharide+ceramide, etc.)

Readings - Lecture 2 Lehninger 5th edition 2008

Chapter 10, pp 343-345, 349-357