Cellular Fatty Acid-Binding Proteins II: Proceedings of the 2nd International Workshop on Fatty Acid-Binding Proteins, Maastricht, August 31 and September 1, 1992

201 Pages·1993·1.53 MB·English

by  Jan F. C. Glatz

Cellular Fatty Acid-Binding Proteins II
Developments in Molecular and Cellular Biochemistry
Series Editor: Naranjan S. Dhalla, Ph.D., FACC
1. V.A. Najjar (ed.): Biological Effects ofGlutamic Acid and Its Derivatives. 1981 ISBN 90-6193-841-4
2. V.A. Najjar (ed.): Immunologically Active Peptides. 1981 ISBN 90-6193-842-2
3. V.A. Najjar (ed.): Enzyme Induction and Modulation. 1983 ISBN 0-89838-583-0
4. V.A. Najjar and L. Lorand (eds.): Transglutaminase. 1984 ISBN 0-89838-593-8
5. G.J. van der Vusse (ed.): Lipid Metabolism in Normoxic and Ischemic Heart. 1989 ISBN 0-7923-0479-9
6. J.F.c. Glatz and G.J. van der Vusse (eds.): Cellular Fatty Acid-Binding Proteins. 1990
ISBN 0-7923-0896-4
7. H.E. Morgan (ed.): Molecular Mechanisms ofCellular Growth. 1991 ISBN 0-7923-1183-3
8. G.J. van der Vusse and H. Stam (eds.): Lipid Metabolism in the Healthy and Diseased Heart. 1992
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10. J.F.c. Glatz and G.J. van der Vusse (eds.): Cellular Fatty-Acid-Binding Proteins II. 1993
ISBN 0-7923-2395-5
Springer Science+Business Media, B.V
Cellular
Fatty Acid-Binding Proteins
Proceedings of the 2nd International Workshop
on Fatty Acid-Binding Proteins, Maastricht,
August 31 and September 1, 1992
edited by
lAN F. C. GLATZ & GER l. VAN DER VUSSE
Department of Physiology
University of Limburg
Maastricht, The Netherlands
Reprinted from Molecular and Cellular Biochemistry, Volume 123, Nos. 1 & 2 (1993)
SPRINGER SCIENCE+BUSINESS, MEDIA, B.V.
Library of Congress Cataloging-in-Publication Data
International Workshop en Fatty ACld-6lndlng Prete,ns (2nd , 1992
Maastr,cht, NetherlandsJ
Cellular fatty aCld-b,nc,ng pretllns II procI.dlngs of the 2nd
International Werkshop on Fatty Ac,d-6Indlng Prote,ns, MaaStricht,
August 31 and September 1, 1992 / edlted by Jan F,C. Olan, Oer J.
van der VUS511.
p. cm. -- (Dev.lop~ents In l1101ecular ano. cellular
b,ochemlstry ; 10l
"Reprlnted from Molecular anO. cellular blochem,stry, voi. 123.
nos. 1 & 2 (1993l."
ISBN 978-1-4613-6353-8 ISBN 978-1-4615-3096-1 (eBook)
DOI 10.1007/978-1-4615-3096-1
1. Fatty aCld-blnding protllns--Congresses. r. Olatz, Jan F. C.
II. Vun •• 13. J. van der. III. Tltl,. IV. Sules, Oevelopments In
molecular and Clliular blochemlstry ; v. 10.
OP552.F37I57 1992
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ISBN 978-1-4613-6353-8
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© 1993 Springer Science+Business Media Dordrecht
Originally published by Kluwer Academic Publishers in 1993
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Molecular and Cellular Biochemistry:
An International Journal for Chemical Biology in Health and Disease
CONTENTS
CELLULAR FArry ACID-BINDING PROTEINS II
Preface
G. Scapin, A.C.M. Young, A. Kromminga, J.H. Veerkamp, J.1. Gordon and J.C. Sacchettini: High resolution X-ray
studies of mammalian intestinal and muscle fatty acid-binding proteins provide an opportunity for defining the
chemical nature of fatty acid: protein interactions 3
D. Lassen, C. Liicke, A. Kromminga, A. Lezius, F. Spener and H. Riiterjans: Solution structure of bovine heart fatty
acid-binding protein (H-FABPe) 15
T. Borchers and F. Spener: Involvement of arginine in the binding of heme and fatty acids to fatty acid-binding protein
from bovine liver 23
KR. Miller and D.P. Cistola: Titration calorimetry as a binding assay for lipid-binding proteins 29
G.E. Gerber, D. Mangroo and B. L. Trigatti: Identification of high affinity membrane-bound fatty acid-binding proteins
using a photoreactive fatty acid 39
J. Storch: Diversity of fatty acid-binding protein structure and function: studies with fluorescent ligands 45
S. Mandrup, P. H. Andreasen, J. Knudsen and K Kristiansen: Genome organization and expression of the rat ACBP
gene family 55
J.M. Stephens, M. Butts, R Stone, P.H. Pekala and D.A. Bernlohr: Regulation of transcription factor mRNA accumula-
tion during 3T3-Ll preadipocyte differentiation by antagonists of adipogenesis 63
F. Schroeder, J.R Jefferson, D. Powell, S. Incerpi, J.K Woodford, S.M.Colles, S. Myers-Payne, T. Emge, T. Hubbell,
D. Moncecchi, D.R Prows and e.E. Heyliger: Expression of rat L-FABP in mouse fibroblasts: role in fat absorption 73
A. Mallordy, P. Besnard and H. Carlier: Research of an in vitro model to study the expression of fatty acid-binding
proteins in the small intestine 85
RM. Kaikaus, W.K Chan, P.R Ortiz de Montellano and N.M. Bass: Mechanisms of regulation of liver fatty acid-
binding protein 93
J.H. Veerkamp and H.T.B. Van Moerkerk: Fatty acid-binding protein and its relation to fatty acid oxidation 101
A Garnier, e. Poizat, C. Keriel, P. Cuchet, M. M. Vork, Y.F. De Jong and J.F.e. Glatz: Modulation of fatty acid-
binding protein content of adult rat heart in response to chronic changes in plasma lipid levels 107
S. Iseki, O. Amano, T. Kanda, H. Fujii and T. Ono: Expression and localization of intestinal 15 kDa protein in the rat 113
P. A Sellner: Retinal FABP principally localizes to neurons and not to glial cells 121
J. Knudsen, S. Mandrup, J.T. Rasmussen, P.H. Andreasen, F. Poulsen and K. Kristiansen: The function of acyl-CoA-
binding protein (ACBP)IDiazepam binding inhibitor (DBI) 129
AK. Dutta-Roy, M.J. Gordon, D.J. Leishman, BJ. Paterson, G.G. Duthie and W.P.T. James: Purification and partial
characterisation of an a-tocopherol-binding protein from rabbit heart cytosol 139
DJ. Van der Horst, J.M. Van Doorn, P.e.e.M. Passier, M.M. Vork and J.F.e. Glatz: Role of fatty acid-binding protein
in lipid metabolism of insect flight muscle 145
N.H. Haunerland, X. Chen, P. Andolfatto, J.M. Chisholm and Z. Wang: Developmental changes of FABP concentra-
tion, expression, and intracellular distribution in locust flight muscle 153
E.A. Meijer, S.e. de Vries, P. Sterk, D.WJ. Gadella Jr., KW.A. Wirtz and T. Hendriks: Characterization of the non-
specific lipid transfer protein EP2 from carrot (Daucus carota L.) 159
J.F.e. Glatz, M.M. York and GJ. van der Vusse: Significance of cytoplasmic fatty acid-binding protein for the
ischemic heart 167
M.M. Vork, J.F.e. Glatz and G.J. van der Vusse: Release of fatty acid-binding protein and long chain fatty acids from
isolated rat heart after ischemia and subsequent calcium paradox 175
P.G.A Voiders, M.M. York, J.F.C. Glatz and J.F.M. Smits: Fatty acid-binding proteinuria diagnoses myocardial
infarction in the rat 185
N.M. Bass: Cellular binding proteins for fatty acids and retinoids: similar or specialized functions? 191
Index 203
Molecular and Cellular Biochemistry 123: 1, 1993.
© 1993 Kluwer Academic Publishers.
Preface
Twenty years have elapsed since cytoplasmic proteins exhibiting high-affinity binding of long-chain fatty acids were
first identified (Ockner et at., Science 177: 56-58,1972). These cellular fatty acid-binding proteins (FABPs) are now well
established to comprise a ligand-defined group of macromolecules belonging to a family of cytoplasmic lipid binding
proteins. Unique features ofthe FABPs are the existence of distinct types ofFABP and that these are found in a variety
of tissues in remarkable abundance, with some cells expressing more than one type. The physiological significance of
the FABPs has only partly been elucidated. By increasing the cytoplasmic solubilization of fatty acids, the cellular
FABPs are considered to function primarily in intracellular fatty acid transport, but may also be assigned important
regulatory roles in cellular lipid homeostasis as well as in the modulation of cell growth and differentiation.
The broad interests in cellular FABPs had led to the organization of the 1st International Workshop on Fatty Acid
Binding Proteins, held in Maastricht, the Netherlands, in 1989. The proceedings of this workshop have been published
in Molecular and Cellular Biochemistry, Volume 98, 1990. Prompted by the success of the first meeting, the 2nd In
ternational Workshop on Fatty Acid-Binding Proteins, which was held again in Maastricht, on August 31 and Septem
ber 1, 1992, brought together scientific specialists in the field of FABP research for two days of intensive and fruitful
discussion. This focussed issue is a collection of selected papers from this conference, and thus provides the state-of
the-art knowledge of cellular FABPs. The contributors to this issue represent pioneering as well as new investigators,
and also reflect the multidisciplinary nature of research in this exciting and rapidly progressing field. We hope that the
present report will accelerate our understanding of the significance of these proteins for the functioning of the celL
Maastricht, December 1992
Jan F.e. Glatz and Ger J. van der Vusse
Department of Physiology
Cardiovascular Research Institute Maastricht
University of Limburg, Maastricht,
The Netherlands
Molecular and Cellular Biochemistry U3: 3-13, 1993.
© 1993 Kluwer Academic Publishers.
High resolution X-ray studies of mammalian
intestinal and muscle fatty acid-binding proteins
provide an opportunity for defining the
chemical nature of fatty acid: protein
interactions
Giovanna Scapin,! Aideen C.M. Young,l Arno Kromminga,l
Jacques H. Veerkamp,3 Jeffrey I. Gordon2 and James C.
Sacchettini1
Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY 10461,2 Department of Molecular
I
Biology and Pharmacology, Washington University School of Medicine, St. Louis, Mo. 63110, and Department of
3
Biochemistry, University of Nijmegen, Nijmegen, The Netherlands
Abstract
The structure of E. coli-derived rat intestinal fatty acid-binding protein has recently been refined to 1.2 A without
bound fatty acid and to 2.0 A and 1.75 A with bound hexadecanoate (palmitate) and 9Z-octadecenoate (oleate), re
spectively. The structure of E. coli-derived human muscle fatty acid-binding protein has also been solved to 2.1 A with a
C16 bacterial fatty acid. Both proteins contain 10 anti-parallel p-strands in a + 1, + 1, + 1. .. motif. The strands are
arranged in two p-pleated sheets that are orthogonally oriented. In each case, the fatty acid is enclosed by the p-sheets
and is bound to the proteins by feeble forces. These feeble forces consist of (i) a hydrogen bonding network between
the fatty acid's carboxylate group, ordered solvent, and side chains of polar/ionizable amino acid residues; (ii) van der
Waals contacts between the methylene chain of the fatty acid and the side chain atoms of hydrophobic and aromatic
residues; (iii) van der Waals interactions between the m-terminal methyl and the component methenyls of the phenyl
side chain of a Phe which serves as an adjustable terminal sensor situated over a surface opening or portal connecting
interior and exterior solvent; and (iv) van der Waals contacts between methylenes of the alkyl chain and oxygens of
ordered waters that have been located inside the binding cavity. These waters are positioned over one face of the ligand
and are held in place by hydrogen bonding with one another and with the side chains of protein's polar and ionizable
residues. Binding of the fatty aeid ligand is associated with minimal adjustments of the positions of main chain or side
chain atoms. However, acquisition of ligand is associated with removal of ordered interior solvent suggesting that the
free energy of dehydration of the binding site may be as important for the energy of the binding reaction as the free
energy of stabilization of the fatty aeid : protein complex. (Mol Cell Bioehem U3: 3-13, 1993)
Key words: fatty acid-protein interactions, X-ray crystallography
Address for offprints: J.c. Sacchettini, Department of Biochemistry, Albert Einstein College of Medicine. 1300 Morris Park Ave, Bronx, New York
10461, USA
4
Introduction rna Chemical Company, St. Louis, MO) at 3]0 C [1]. The
delipidated protein (apo-I-FABP) was concentrated to
15-30 mg/ml (1-2 mM) in 100 mM piperazine-N,N'bis[2-
The structures of several members of a family of ver
ethanesulfonic acid] (PIPES), pH 7.1, 0.05% sodium
tebrate intracellular lipid binding proteins have been
azide and reacted with a two fold molar excess of pure
solved during the last 3 years using x-ray crystallograph
fatty acid for 15 min at 37° C. Crystals of apo-I-FABP
ic methods. These include rat intestinal fatty acid-bind
or of the binary I-FABP: fatty acid complex (binding
ing protein (I-FABP, [1-4]) bovine heart fatty acid-bind
ing protein (H-FABP, [5]), the P2 protein of bovine pe stoichiometry = 1 : 1) were produced using the free in
terface diffusion method [9]. This method involves plac
ripheral nerve myelin [6], chicken liver fatty acid-bind
ing protein (L-FABP, [4]), and mouse adipocyte lipid ing of250 ~l of the concentrated I-FABP solution over a
solution of polyethyleneglycol (24-28% (w/v) prepared
binding protein (ALBP, [7]). All of these proteins con
in 100 mM PIPES, pH 7.1, 0.05% sodium azide; average
tain 10 anti-parallel ~-strands and two short a-helices.
molecular weight of the PEG = 4000) and incubating the
The overall appearance of the structures resembles that
material in a Teflon-stoppered glass vial at 19° C. Crys
of a clam shell: two nearly orthogonal ~-sheets envelop
tals could be grown to 2.0 x 1.0 x 1.0 mm within 2 weeks.
the bound fatty acid which is located in a large, interior,
Table 1 lists the properties of the crystals of apo-I-FABP,
solvent-filled cavity. Comparisons of the structures of
I-FABP: palmitate and I-FABP: oleate.
these proteins and the conformations of their bound fat
ty acids have provided an opportunity for describing the
atomic details of fatty acid protein interactions. In this
Purification and crystallization ofE. coli-derived human
report, we compare the recently refined high resolution
muscle fatty acid-binding protein
structures of E. coli-derived rat intestinal fatty acid
binding protein, with or without a homogeneous pop
The methods used to produce M-FABP in E. coli and
ulation of bound palmitate (hexadecanoic acid; CI6 : 0)
then purify it to apparent homogeneity have been de
or oleate (9Z-octadecenoic acid: CI8 : I), and the struc
scribed in recent papers [10, 11]. The procedures employ
ture of E. coli-derived human muscle fatty acid-binding
ed for preparing apo-M-FABP and M-FABP with
protein containing a bound C16 bacterial fatty acid.
bound fatty acids were similar to those described above
for the homologous intestinal fatty acid-binding protein.
Experimental procedures Crystals of apo-M-FABP, M-FABP with a popUlation of
bound bacterial fatty acids, and M-FABP with a homo
geneous population of fatty acid ligand were obtained
Purification and crystallization of E. coli-derived rat
intestinal fatty acid-binding protein using the hanging drop vapor diffusion method. Five ~l
of protein (25 mg/mL prepared in 20 mM PIPES,
The host-vector system used to produce rat I-FABP in pH 7.1,0.05% sodium azide) were mixed with 5 ~l of
30% polyethyleneglycol4000 (in 20 mM PIPES, pH 7.1,
E. coli and the methods employed to purify the protein
0.05% azide) on a silanized glass coverslip. The cover
from bacterial lysates have been described in earlier
slip was then inverted, placed over a well containing 1 ml
publications [4, 8]. Bound bacterial fatty acids were re
of the same PEG solution, and stored at 19° C. Crystals
moved from the purified protein by passage through hy
appeared and reached a maximum size of 0.7 x 0.5 x
droxyalkoxypropyl dextran (Lipidex 1000, type VI, Sig-
Table 1. Characterization of crystals of apo-I-FABP and I-FABP: fatty acid complexes
Complex Space group Molecules! a b c IX ~ Y Resolution
e)
Asymmetric unit (A) (A) (A) (0) (0) (A)
Palmitate (I) P2, I 36.9 56.8 31.8 90.0 114.0 90.0 2.0
Palmitate (II) P2, 2 68.1 56.2 37.7 90.0 104.0 90.0 1.9
Oleate P2, 2 68.2 56.7 37.8 90.0 104.5 90.0 1.8
Apo (I) P2, 1 36.0 56.6 31.6 90.0 113.1 90.0 1.9
Apo (II) P2, 1 35.8 51.4 31.3 90.0 91.4 90.0 0.9
5
Table 2. Crystallographic properties of complexes of M-FABP with different ligands
Complex Space group a b c a 13 y resolution
(A) (A) (A) n CO) (0) (A)
Native P2,2,2, 35.4 56.7 72.7 90.0 90.0 90.0 2.1
Stearate 34.56 55.26 71.17 90.0 90.0 90.0 1.3
Oleate 34.67 55.48 71.86 90.0 90.0 90.0 1.3
Arachidonate 34.39 55.49 71.30 90.0 90.0 90.0 1.45
Elaidate 34.63 55.24 71.41 90.0 90.0 90.0 1.4
0.5 mm within 2 weeks. The characteristics of crystals of FABP: palmitate, I-FABP: oleate and apo-I-FABP are
M-FABP containing a bound bacterial fatty acid or a described in references 1-4. Panels A and B of Fig. 1
specific CIS or C20 fatty acid are described in Table 2. show the backbone structures and the location of the
bound fatty acid for the two holo-proteins. Panel C
shows the Ca trace for the apo-protein and the local
Data collection and model refinement ization of24 ordered water molecules that have been lo
cated in the binding cavity. The protein contains ten an
Details of data collection, data analyses, and refinement ti-parallel ~-strands (~A-~J), arranged into two ortho
procedures can be found in our earlier papers [1-4, 11]. gonal ~-pleated sheets. The first J3-sheet is composed of
The structures of the apo- and holo-proteins were re strands ~A through the first five residues of ~F (Ala! ~
fined to high resolution using a combination of least GilD). The second sheet contains the remaining 5 resid
square methods [12], energy refinement and molecular ues of ~F through ~J (ThrR! ~ Glu131). Tyr!4 ~ Lys20 and
dynamics [131 plus manual model building. The statistics Val25 ~ Gly3! form the two short a-helices (aI and all)
after refinement of I-FABP and M-FABP, with and of I-FABP. They are connected by a loop (Mee! ~
without bound fatty acid, are described in Table 3. Asn24) and are inserted between the first two ~-strands.
The helices are both right handed and are maintained by
typical n to n + 4 main chain hydrogen bonds.
Results and discussion I-FABP contains 42 ionizable, 26 polar and 46 hydro
phobic residues. The remaining residues are glycines.
The three dimensional structure of I-FABP with and The side chains of 33 ionizable, 19 polar, and 12 hydro
without bound fatty acids phobic residues form a relatively uniform shell around
the molecule. Many hydrogen bonds occur between po
The high resolution structures of E. coli-derived rat 1- lar and ionizable residues located at the surface of the
Table 3. Statistics after refinement I-FABP and M-FABP
Protein Apo-I-FABP I-FABP: I-FABP: oleate Native M-FABP M-FABP: M-FABP: M-FABP:
palmitate stearate' oleate' elaidate'
Resolution
(A) 1.19 2.0 1.75 2.1 1.3 1.4 1.4
R-faetor % 16.9 17.8 18.3 19.5 17.8 18.3 18.1
Protein atoms 1096 1096 2192' 1030 1030 1030 1030
Solvent atoms 237 60 340' 56 167 152 151
Fatty acid atoms 18 40' 18 20 20 20
RMS deviation from ideal stereochemistry
Bond
distance (A) 0.009 0.012 0.007 0.013 0.010 0.009 0.009
angle (0) 2.85 2.40 1.90 2.70 2.12 2.72 2.21
, Manuscripts in preparation. ' 2 molecules/asymmetric unit.