Table Of Contentiv Preface
also make it interesting to read. It is our wish that this handbook will provide the expert
with an opportunity to become acquainted with complementary information, while it will
offer the beginner a concise survey of the entire field of phospholipid research and a key to
relevant specialized knowledge.
Contents
Gregor Cevc
Preface iii
Contributors ix
Part I: General and Chemical Properties
1. Structure and Nomenclature 1
John R. Silvius
2. Occurrence and Response to Environmental Stresses in Nonmammalian
Organisms 23
S. L. Neidleman
3. Isolation and Analysis of Phospholipids and Phospholipid Mixtures 39
K.-H. Gober, B. R. Giinther, E. M. Liinebach, G. Repplinger, and M. Wiedemann
4. Phospholipid Biosynthesis 65
Kenneth J. Longmuir
5. Chemical Preparation of Sphingosine and Sphingolipids: A Review of Enantioselec-
tive Syntheses 97
Hoe-Sup Byun and Robert Bittman
6. Chemical Preparation of Glycerolipids: A Review of Recent Syntheses 141
Robert Bittman
7. Polymerizable Phospholipids 233
Alok Singh and Joel M. Schnur
8. Coupling and Labeling of Phospholipids 293
Vladimir P. Torchilin and Alexander L. Klibanov
Contents Contents
9. Chemical Stability 323 24. Immunologic Properties and Activities of Phospholipids 817
R. Evstigneeva William E. Fogler
10. Physical Stability on Long-Term Storage 335 25. Phospholipids in Diagnosis 833
Daan J. A. Crommelin, Herre Talsma, Mustafa Grit, and Nicolaas J. Zuidam R. Andrew Badley, Paul J. Davis, and D. M. Tolley
Part II: Physical and Structural Properties 26. Biological and Biotechnological Applications of Phospholipids 855
Roger R. C. New
11. Physical Characterization 35 1
Gregor Cevc and John M. Seddon 27. Medical Applications of Phospholipids 879
Yukihiro Namba
12. Lipid Polymorphism: Structure and Stability of Lyotropic Mesophases of
Phospholipids 403 28. Phospholipids in Disease 895
John M. Seddon and Gregor Cevc C. M. Gupta
13. Dynamic Properties 455 Appendix A: Structural Parameters of Phospholipids 909
Alfred Blume John M. Seddon
14. Ionization and Ion Binding 51 1 Appendix B: Thermodynamic Parameters of Phospholipids 939
Suren A. Tatulian Gregor Cevc
15. Phospholipid Hydration 553 Appendix C: Mechanical, Solubility, and Related Parameters of Phospholipids 957
Thomas J. Mclntosh and Alan D. Magid Gregor Cevc
16. Phospholipid Monolayers 579 Index 963
Helmuth Mohwald
17. Phospholipid Vesicles 603
Helmut Hauser
18. Solute Transport Across Bilayers 639
Gregor Cevc
19. Intermembrane and Transbilayer Transfer of Phospholipids 663
J. Wylie Nichols
20. Magnetic Resonance Studies of Phospholipid-Protein Interactions in Bilayers 687
Anthony Watts
Part Ill: Biological Aspects
21. Biological Distribution 745
Mark A. Yorek
22. Phospholipid Metabolism in Animal Cells 777
Gerrit L. Scherphof
23. Toxicity and Systemic Effects of Phospholipids 801
Theresa M. Allen
Contributors
Theresa M. Allen Department of Pharmacology, University of Alberta, Edmonton,
Alberta, Canada
R. Andrew Badley Immunology Section, Unilever Research, Bedford, England
Robert Bittman Department of Chemistry and Biochemistry, Queens College of The
City University of New York, Flushing, New York
Alfred Blume Department of Chemistry, University of Kaiserslautern, Kaiserslautern,
Germany
Hoe-Sup Byun Department of Chemistry and Biochemistry, Queens College of The
City University of New York, Flushing, New York
Gregor Cevc Medical Biophysics Laboratory, Technical University of Munich,
Munich, Germany
Daan J. A. Crsmmelin Department of Pharmaceutics, University of Utrecht, Utrecht,
The Netherlands
Paul J. Davis Immunology Section, Unilever Research, Bedford, England
R. Evstigneeva Lomonosov Institute of Fine Chemical Technology, Moscow, Russia
William E. Fogler Laboratory of Experimental Immunology, Biological Response
Modifiers Program, DCT, NCI-FCRDC, Frederick, Maryland
K.-H. Gober Analytical Development, RhGne-Poulenc Rorer GmbH, Koln, Germany
Mustafa Grit Department of Pharmaceutics, University of Utrecht, Utrecht, The
Netherlands
x Contributors
Contributors xi
B. R. Giinther Process Development, RhBne-Poulenc Rorer GmbH, Koln, Germany
Alok Singh Center for Bio/Molecular Science and Engineering, Naval Research Lab-
oratory, Washington, D.C.
C. M. Gupta Institute of Microbial Technology, Chandigarh, U. T., India
Herre Talsma Department of Pharmaceutics, University of Utrecht, Utrecht, The
Helmut Hauser Department of Biochemistry, ETH Zurich, Zurich, Switzerland
Netherlands
Alexander L. Klibanov Department of Pharmacology, University of Pittsburgh
Suren A. Tatulian* Institute of Cytology, Academy of Sciences of the Russian
School of Medicine, Pittsburgh, Pennsylvania
Federation, St. Petersburg, Russia
Kenneth J. Longmuir Department of Physiology and Biophysics, College of Medi-
I). M. Talky Department of Chemistry, Unipath Ltd., Bedford, England
cine, University of California, Irvine, California
Vladimir P. Torchil in Center for Imaging and Pharmaceutical Research, Massachu-
E. M. Lunebach Analytical Development, RhBne-Poulenc Rorer GmbH, Koln, Ger-
setts General Hospital-East, Charlestown, Massachusetts
many
Anthony Watts Department of Biochemistry, Oxford University, Oxford, England
Alan D. Magid Department of Cell Biology, Duke University Medical Center,
Durham, North Carolina
M. Wiedemann Process Development, RhBne-Poulenc Rorer GmbH, Koln, Germany
Thomas J. Mclntosh Department of Cell Biology, Duke University Medical Center,
Mark A. Yorek Department of Internal Medicine, Veterans Administration Medical
Durham, North Carolina
Center, Iowa City, Iowa
Helmuth Mohwald Institute of Physical Chemistry, Johannes-Gutenberg-University
Nicolaas J. Zuidam Department of Pharmaceutics, University of Utrecht, Utrecht,
of Mainz, Mainz, Germany
The Netherlands
Yukihiro Namba Lipid Project, Nippon Fine Chemical Co., Ltd., Hyogo, Japan
S. L. Neidleman Biosource Genetics Corporation, Vacaville, California
Roger R. C. New* Cortecs Ltd., London, England
J. Wylie Nichols Department of Physiology, Emory University School of Medicine,
Atlanta, Georgia
G. Repplinger Analytical Development, RhBne-Poulenc Rorer GmbH, Koln, Ger-
many
Gerrit L. Scherphof Department of Physiological Chemistry, Groningen Institute for
Drug Studies, University of Groningen, Groningen, The Netherlands
Joel M. Schnur Center for Bio/Molecular Science and Engineering, Naval Research
Laboratory, Washington, D. C.
John M. Seddon Department of Chemistry, Imperial College, London, England
John R. Silvius Department of Biochemistry, McGill University, Montreal, Quebec,
Canada
*Former affiliation: Biocompatibles Ltd., Middlesex, England
*Current affiliation: Department of Physiology, University of Virginia, Charlottesville, Virginia
I
General and Chemical Properties
An enormous variety of phospholipid structures is found in nature, exhibiting great
diversity in the structures of both the apolar and the polar moieties of the lipid molecules.
While any individual lipid species may be named according to rigorous rules of organic-
chemical nomenclature [I-51, a practical system of nomenclature for biologically occur-
ring lipids must also offer reasonable brevity and simplicity to be generally useful.
Moreover, even a 'purified' lipid preparation that is obtained from a biological source
may be homogeneous with respect to one structural feature (e.g., the polar portions of the
lipid molecules) but highly heterogeneous with respect to another (e.g., their hydrocarbon
chains). A useful system of nomenclature for lipids from natural sources must allow
for this fact by permitting certain structural features of the lipids in a given preparation
to be described in a generic manner while other features of the structure are specified
entirely.
Three general types of nomenclature for lipids will be described in this chapter. In
what will be referred to as the 'fully systematic' or 'formal' system of nomenclature, all
acyl and alk(en)yl residues are fully specified, using their systematic (IUPAC) designa-
tions [I], and all other structural units in the lipid molecule (polyols, monosaccharide
units, amino acids, phospho- moieties, etc.) are specified individually and in full, using
the nomenclature recommended in the IUPAC-IUB proposals for the Nomenclature of
Lipids [4], Nomenclature of Phosphorus-Containing Compounds of Biological Im-
portance [3] and Prenol Nomenclature [5]. In systems of nomenclature referred to below
as 'trivial,' details of stereochemistry are often absent or implicit, and major portions of
the molecule may be named as simple units (e.g., as a 'phosphatidyl' group) or generical-
ly. Finally, shorthand systems of nomenclature have been developed, using simple
alphabetical or numerical symbols to allow concise specification of the structures of even
complex lipid molecules (e.g., glycolipids) or to focus on particular details of molecular
structure, such as the fatty acyl composition of a natural lipid preparation.
It is common in practice to define completely the structures of both the apolar and the
polar portions of pure synthetic phospholipids, which constitute single molecular species.
2 Silvius Structure and Nomenclature
By contrast, it is common to specify in detail only the polar portions of 'pure' lipid
fractions from natural sources while designating the apolar portions, which are normally
heterogeneous in composition, in a generic manner. Accordingly, this chapter will first
describe the systems of nomenclature used to define the structures of the apolar and HO-~$-H 'sn-glycerol'
'backbone' portions of lipid molecules. The following sections describe systems of
nomenclature that are appropriate to describe either individual phospholipid molecular
species or phospholipid fractions that are uniform in some structural features but heteroge-
neous in others.
II. APOLAR AND 'BACKBONE' MOIETIES OF
PHOSPHOLIPIDS - .
1
Almost all biologically occurring phospholipids are constructed from two combinations of
apolar and 'backbone' moieties: a glycerol (or other polyol) moiety substituted with one or OH
4 -
two acyl or alkyl chains; or an N-acylated sphingoid base (i.e., a ceramide).
Sphingenine ( Sphingosine ) NH2
A. Structure and Nomenclature of Acyl/Alkylated
Glycerol Moieties
While glycerol itself is a symmetrical molecule, its carbon-2 becomes a chiral center when
the 1- and 3-positions are not symmetrically substituted. It is therefore useful to define a
prochiral 'sn-glycerol' (sn = stereospecific numbering [6]) in which the orientation of the
2-hydroxyl group and the numbering of the carbons are as shown in Fig. 1. In virtually all lcosasphingenine
natural phospholipids, excepting certain archaebacterial lipids discussed below, the polar
headgroup is attached to the 3-position of 'sn-glycerol,' in which the configuration of
substituents about C-2 would be designated as R in the (R,S)-system. The 'sn-' conven-
tion is also applied to describe the configuration of other glycerol residues in phospholipid
molecules (e.g . , for the biologically occurring 1-(1' ,2' -diacyl-sn-glycero-3-phospho)-sn-
-
glycerol and its 3-0-amino acyl esters [7]). Glycerol residues that are not either sym-
metrically or stereospecifically substituted are designated using the prefix rac-, as in Sphinganine ( Dihydrosphingosine ) NH2
1-(1' ,2'-diacyl-sn-glycero-3'-phospho)-rac-glycerol,in which the nonacylated glycerol
comprises a mixture of 1- and 3-substituted 'sn-glycerol' residues.
1. Acyl Residues
Acyl chains are named formally using standard IUPAC rules of nomenclature for organic
compounds [I], although certain features of the IUPAC system, notably the use of the
(E,Z)-convention to designate the configuration about the double bond, have not received
widespread use in lipid nomenclature. The systematic name of an acyl residue is derived 4-D-Hydroxysphinganine ( Phytosphingosine )
from that of the corresponding fatty acid by replacing the suffix '-oic acid-' by '-oyl,' as in
hexadecanoyl or cis, cis-9,12-octadecadienoyl. Figure 1 Structures of sn-glycerol and of the major sphingoid bases.
Most of the fatty acids found in natural phospholipids also have one or more trivial
names (Table 1; for more extensive compilations see [8,9]). To designate the name of an
acyl residue using the trivial nomenclature, the suffix '-ic acid' in the common name of
the parent fatty acid is replaced by the suffix '-oyl,' in analogy to the designation of acyl
residues in the IUPAC system. This rule can lead to confusion in certain cases, however,
notably for decanoyl chains, which it designated as 'caproyl' (from capric acid), could be
mistaken for a residue of esterified hexanoic (caproic) acid.
Several common systems also exist for the shorthand designation of fatty acyl chains.
The first, which is used in an IUPAC-IUB-recommended system for the shorthand
Silvius Structure and Nomenclature 7
alcohols linked to the diacylglycerophospho- moiety* are normally designated by their
common names (e.g., ethanolamine rather than 2-aminoethanol) unless the alcohol
substituent is a rare or unnatural compound. In this system, for example, dipalmitoylphos-
phatidylcholine (Fig. 2a) is designated as 1,2-dihexadecanoyl-sn-glycero-3-phospho-
choline or, less commonly in practice, 1,2-dihexadecanoyl-sn-glycero(3)phosphocholine
PI.
Various trivial nomenclatures exist for diacylglycerophospholipids, of which the
most useful (and precise) defines such compounds as derived from a phosphatidic acid,
i.e., a diacylglycerophosphate. Lipids in which phosphatidic acids are coupled to alcohols
in phosphodiester linkage are then named as phosphatidyl alcohols (e.g., phosphati-
dylcholine). The term 'phosphatidyl' used without qualification generally implies
a 1,2-diacyl-sn-glycero-3-phospho-m oiety. It has been recommended [4] that this
and particularly other, less common diacylglycerophospho- residues be named as 'x-sn-
phosphatidyl' residues, where x is the position of the phospho- group on the sn-glycero-
backbone. However, the designation '(3)-phosphatidyl(alcoho1)' is seldom found in
common use. While the strict definition of a phosphatidyl residue is a diacyl-
glycerophospho- moiety, the use of the term is commonly (if incorrectly) extended to en-
compass mixtures of diacyl- and alkylacylglycerophospholipids in cases where the two
are not resolved (e.g., in most chromatographic procedures). It should be noted that
9v-
a 'phosphatidyl' residue actually comprises the unit (diacylglycero1)-0- 'Phospha-
0
tidylethylamine,' for example, thus designates a phosphonolipid (see below) and not
phosphatidylethanolamine.
Most older systems of nomenclature used for phospholipids offer no special advan-
tages over those described above, and their use is not recommended. Terms such as
'lecithin' and 'cephalin' were originally applied to describe crude lipid fractions from
particular tissues and are inappropriate for use to describe pure lipid species. The older
designation of the 3-, 2- and 1-positions of 'sn-glycerol' as the a-, P- and y-positions,
with phospholipids from eubacteria and eukaryotes defined as derivatives of 'L-a'-phos-
phatidic acid, should likewise be abandoned in favor of the systems described above.
As noted above, a shorthand nomenclature has been proposed for glycerophospho-
lipids as well as other complex lipids [3,4].T he complete shorthand designation for a
1,2-diacyl-sn-glycero-3-phosphate-derivelidp id consists of shorthand designations (see
Table 1) for the acyl chains at the I- and 2-positions (or a single abbreviation with a
subscript '2' if the two chains are the same), followed successively by the abbreviation
Gro and the shorthand designation of the phosphodiester-linked alcohol (see Table 2). The
phosphatidyl moiety may also be denoted by the generic abbreviation Ptd. Using these
conventions, dipalmitoyl phosphatidylcholine becomes Pam2GroPCho (or, in a variant
usage, Pam2PtdCho) and phosphatidylserine (with a mixture of fatty acyl chains) becomes
Figure 2 Structures of representative phospholipids with configurations characteristic of eukaryo- *The IUPAC recommendations for nomenclature of phosphorus-containing compounds [3] suggest that
tic and eubacterial phospholipids. (a) 1,2-dihexadecanoyl-sn-glycero-3-phosphocholineo r L- a- phosphodiesters be designated by inserting the infix 'phospho' (as a contraction of 'phosphinico') between the
names of the two coupled alcohols, and that phosphomonoesters be designated using the prefix 'phospho' (as a
dipalmitoylphosphatidylcholine. (b) 1-hexadecyl-2-acetyl-sn-glycero-3-phosphochoine (platelet
contraction of 'phosphono'), in cases where it is not necessary or possible to specify the other substituents on the
activating factor). (c) A 1-(alk-1 '-eny1)-2-acyl-sn-glycero-3-phospho(alcoho1a)l so named as an
phosphoryl residue (e.g., hydrogen). Phosphomonoester compounds may also be named as alcohol phosphates
(alcohol) plasmalogen. The most common moieties X in naturally occurring phospholipids are
in such cases. It is recommended that the term 'phosphoryl' be reserved for cases in which all three substituents
ethanolamine and choline. (d) A 1-(1 '-glyceroalkyl)-2-acyI-sn-glycero-3-phosphoethanolamine, ,o
,
found in membranes of various Clostridium species. (e) 2-hexadecanoyl-sn-glycero-3- on the O=P-0 center are specified. The commonly used terminologies 'glycerophosphoryl(alcohol),' 'di-
phosphocholine, also named as 2-palmitoyl-1-lyso-phosphatidylcholine. 'n -
acylglycerophosphoryl(alcohol),' etc. are thus incorrect
