letters

Structure of human a NH2

NH2

monoamine oxidase B,
a drug target for the BENZYLAMINE PHENETHYLAMINE

treatment of neurological N

disorders
© 2002 Nature Publishing Group http://structbio.nature.com

N CH3 HO

Claudia Binda1, Paige Newton-Vinson2, NH2

MPTP SEROTONIN
Frantisek Hubálek2, Dale E. Edmondson2
and Andrea Mattevi1 OH
H
HO NH2 HO N
1Department of Genetics and Microbiology, University of Pavia, Pavia, Italy. CH3
2Department of Biochemistry, Emory University School of Medicine, Atlanta,
HO HO
Georgia, USA.
DOPAMINE EPINEPHRINE (adrenaline)
Published online: 26 November 2001, DOI: 10.1038/nsb732

Monoamine oxidase B (MAO B) is a mitochondrial outer- b

membrane flavoenzyme that is a well-known target for anti-
depressant and neuroprotective drugs. We determined the
structure of the human enzyme to 3 Å resolution. The
enzyme binds to the membrane through a C-terminal trans-
membrane helix and apolar loops located at various posi-
tions in the sequence. The electron density shows that
pargyline, an analog of the clinically used MAO B inhibitor,
deprenyl, binds covalently to the flavin N5 atom. The active
site of MAO B consists of a 420 Å3-hydrophobic substrate
cavity interconnected to an entrance cavity of 290 Å3. The
recognition site for the substrate amino group is an aromat-
ic cage formed by Tyr 398 and Tyr 435. The structure pro-
vides a framework for probing the catalytic mechanism,
understanding the differences between the B- and
A-monoamine oxidase isoforms and designing specific
inhibitors.
Monoamine oxidase B (MAO B) is one of two flavin-depen-
dent isozymes (the other being MAO A) that function in the
oxidative deamination of neurotransmitters and exogenous aryl-
alkylamines (Fig.1a). The electron acceptor for this reaction is
O2, which is converted to hydrogen peroxide by either enzyme
(Fig. 1b,c). Mammalian MAOs are bound to the outer mito-
chondrial membrane through a C-terminal transmembrane
polypeptide segment1. These oxidases have long been a pharma-
cological focus because both reversible and irreversible
inhibitors of MAO A and B have been used clinically in the treat-
ment of neurological disorders. The MAO B inhibitor, deprenyl,
is administered to potentiate L-dopa therapy in the treatment of
Parkinson’s patients as well as to provide neuroprotective effects
in patients exhibiting pre-Parkinson’s Syndrome2. The level of
enzyme expressed is dependent on tissue type and is elevated
three-fold in aged human neuronal tissue3. Recent studies have
demonstrated that elevated MAO B levels induce apoptosis in
neuronal4 and kidney cells5. Increased levels of MAO B have also
been demonstrated in plaque-associated astrocytes of brains
from Alzheimer’s patients6; MAO B inhibitors are currently in
clinical trials for treatment of this disease. This enzyme may also
be important in pathological processes resulting from exposure
to various xenobiological compounds. The first example of a
chemically-induced Parkinson’s Syndrome involves the bioacti-
vation of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP,
an impurity in synthetic heroin) by MAO B7 (Fig. 1a). Recent
NH2 NH2+ O
SUBSTRATE (S) IMINE ALDEHYDE
c E.FADox + S E.FADox-S E.FADred-Imine
O2
Imine
Hydrolysis
E.FADox-Imine
H2O2
Aldehyde + NH4+
Fig. 1 Substrate specificity and catalysis of MAOs. a, Examples of human
MAO substrates. MAO A and B have distinct but partly overlapping
specificities. Benzylamine, phenethylamine and 1-methyl-4-phenyl-
1,2,3,6-tetrahydropyridine (MPTP) are good substrates for MAO B (but
are oxidized at a slower rate by MAO A), whereas serotonin is a specific
substrate for MAO A. Dopamine and epinephrine are oxidized at similar
catalytic efficiencies by both enzymes. b, The oxidation of benzylamine
and the reaction product. c, Scheme for the overall oxidative deamina-
tion reaction catalyzed by MAOs. Oxidation of the amine substrate leads
to the reduction of FAD. The prosthetic group is reoxidized by molecular
oxygen to generate hydrogen peroxide. The imine product is hydrolyzed
in a nonenzymatic process.
studies have also shown that components in tobacco smoke
inhibit MAO B, suggesting a contribution to the addictive prop-
erties of tobacco use8.
From this brief description, MAO B (and MAO A) is clearly
involved in a large number of disease states, affecting large seg-
ments of the aging population in particular. Although consider-
able progress has been achieved in the development of clinically
useful MAO B inhibitors, elucidation of the three-dimensional
structure of the enzyme would facilitate further improvements
in drug design. Here we report the first crystal structure of a
flavin-dependent monoamine oxidase: human MAO B. The
structure was solved by the single isomorphous replacement
method combined with multicrystal 12-fold averaging (Fig. 2).
These results reveal the architecture of the catalytic center and of
sites on the protein that are important for its binding to the
outer membrane of the mitochondrion.

22 nature structural biology • volume 9 number 1 • january 2002

 letters

Fig. 2 Stereo view of the experimental electron density map. The map
was obtained by 12-fold averaging over two crystal forms at 3.0 Å reso-
lution. The contour level is 1.0 σ. Carbons are in black, nitrogens in blue,
oxygens in red and sulfurs in yellow. The figure shows the active site with
the flavin ring covalently bound to Sγ of Cys 397 and the pargyline
inhibitor covalently attached to the N5 atom of the FAD cofactor. The
flavin ring is in a bent, twisted conformation, with an angle of 26°
between the planes of the benzenoid and pyrimidine rings.
© 2002 Nature Publishing Group http://structbio.nature.com

Overall structure of human MAO B efficient detergent extraction13. The protein region responsible
The 520 amino acids of MAO B fold into a compact structure for membrane attachment is formed by the C-terminal amino
(Fig. 3a) that exhibits a topology initially found in p-hydroxy-acids 461–520 (ref. 1). Analysis of the MAO B amino acid
benzoate hydroxylase and then observed in several flavopro- sequence with the program TopPred14 predicts that residues
teins9. A search of the Protein Data Bank10 reveals that the closest
489–515 form a transmembrane helix 27 amino acids long,
structural matches of MAO B are L-amino acid oxidase11 and which is well within the range of values observed for transmem-
polyamine oxidase12, with root mean square (r.m.s.) deviations brane helices in membrane proteins of known three-dimension-
of 2.7 Å (420 Cα atoms with 22% sequence identity) and 3.5 Å al structure15. MAO B is expected to contain additional
(404 Cα atoms with 18% sequence identity), respectively. membrane interaction sites, because C-terminal truncation
mutagenesis experiments (ref.16; P.N.-V. and D.E.E., unpub-
The crystal structure shows that the enzyme is dimeric (Fig. 3a),
lished data) show that various deletions of the C-terminal
which is unlikely to be a crystal packing artifact because the dimer
is present in the two crystal forms (orthorhombic and triclinic)residues do not completely abolish the ability of the enzyme to
employed in the structure determination (Table 1; Methods). bind to the membrane.
Indeed, the monomer–monomer interactions are extensive; The crystal structure reveals that the C-terminal residues form
2,095 Å2 of the surface area (15% of monomer accessible surface)an extended polypeptide chain (amino acids 461–488) that tra-
is buried upon dimer formation. Therefore, the dimer form is sug-
verses the monomer surface (Fig. 3a). This extended chain is fol-
gested to be the quaternary assembly present in vivo. lowed by an α-helix that initiates at Val 489, forming the
predicted transmembrane helical segment (Fig. 3a,b). The helix
Structure of the membrane binding region of each monomer protrudes from the basal face of the dimer
MAO B is tightly bound to the outer mitochondrial membrane, (when oriented as in Fig. 3a), with each helical axis approxi-
as evidenced by the need for digestion of phospholipids for its mately parallel to the molecular two-fold axis. This observation

a b c

Fig. 3 Structure of human MAO B. a, Ribbon diagram of the MAO B dimer with the two-fold axis vertical in the plane of the paper. Monomer A is on
the right and monomer B is on the left. The letters ‘N’ and ‘C’ indicate the observed N-terminal (Lys 4 in both monomers) and C-terminal (Thr 500 in
monomer A and Ile 496 in monomer B) amino acids, respectively. Residues 4–460 are in red, and the C-terminal tail (residues 461–500) is in green. The
dimer is anchored to the outer mitochondrial membrane through the C-terminal tails and the neighboring residues of each monomer. The molecu-
lar two-fold axis is perpendicular to the membrane plane, allowing equivalent interactions between each monomer and the membrane. The FAD is
shown in ball-and-stick representation and colored in yellow. The membrane region boundary is indicated by a thick dashed line for illustration pur-
poses and is not intended to define the exact depth of protein insertion into the membrane. b, Close-up view of the membrane binding region in
monomer A. The orientation and the color scheme for the backbone trace are as in (a). To allow visualization of all side chains, the ribbon for the
C-terminal residues (green) has been made semitransparent. Carbons are in black, nitrogens in blue and oxygens in red. The amino-acid sequence of
the C-terminal residues (480–520) is indicated. Residues 489–515 (predicted to form the transmembrane helix) are underlined. Residues 501–520
were not included in the final model because they are not well defined in the electron density. c, Cavities constituting the substrate path from the
protein surface to the flavin in the MAO B monomer. Adjacent to the active site cavity (cyan) is the ‘entrance’ cavity (blue), which is underneath loop
99–112. The FAD is in yellow and the covalent pargyline inhibitor is in black. The color scheme of (a) is used. With respect to (a), the model has been
rotated by 90° around the vertical axis in the plane of the paper.

nature structural biology • volume 9 number 1 • january 2002 23

 letters

a b
© 2002 Nature Publishing Group http://structbio.nature.com

Fig. 4 The substrate binding site of human MAO B. a, Stereo view of the pargyline inhibitor and the residues lining the binding site at the re side of
the flavin. Carbons are in black, nitrogens in blue, oxygens in red and sulfurs in yellow. The inhibitor is outlined by shaded bonds. With respect to
Fig. 3c, the model has been rotated by 90° around an axis approximately orthogonal to the plane of the paper. b, Schematic representation of the
pargyline binding site. MAO B residues that are conserved in human MAO A are indicated by an asterisk. For nonconserved amino acids, the replace-
ment side chains of MAO A are in parentheses. Aromatic side chains are enclosed in ellipsoidal frames. Other residues are in rectangular boxes. The
atoms of the flavin ring are numbered. c, A model for the binding of the substrate, benzylamine, to human MAO B. For illustrative purposes, the
atoms and bonds of the modeled substrate are shown in an increased size. The surface of the solvent inaccessible substrate-binding cavity is semi-
transparent. For clarity, only some of the residues lining the cavity are depicted. Carbons are in cyan, nitrogens in blue and oxygens in red. The ori-
entation is as in (a). The reactive N5 site on the twisted flavin ring is labeled.

suggests that the dimer binds to the membrane with its two-fold
axis perpendicular to the membrane plane, and the C-terminal
helices inserted in the lipid bilayer. In a second helix turn,
Arg 494 is oriented in the proper direction for electrostatic inter-
action with the anionic phospholipid headgroups (Fig. 3b),
which is often observed in transmembrane helices15. Although
the C-terminal helix is predicted to extend to residue 515, the
electron density in this region is interrupted at Thr 500 in
monomer A (Fig. 3b) and at Ile 496 in monomer B. Static and/or
dynamic disorder in this portion of the helix may result from the
absence of the membrane bilayer.
In addition to the C-terminal helical segment, the structure
shows that other protein regions may be involved in membrane
binding (Fig. 3b). In residues 481–488 of the elongated polypep-
tide stretch preceding the C-terminal helix, several hydrophobic
side chains (Phe 481, Leu 482, Leu 486 and Pro 487) point
towards the membrane. At the end of loop 99–112 (Fig. 3b,c),
Pro 109 and Ile 110 are surface-exposed in a position that could
allow interaction with the membrane. The current structural
data do not define the exact depth of protein insertion into the
bilayer. However, the presence of these exposed hydrophobic
side chains suggests that membrane attachment does not solely
involve the C-terminal helix but also additional hydrophobic
patches of the protein surface.
The active site
The electron density map (Fig. 2) shows that pargyline covalent-
ly binds to the N5 atom on the re side of the flavin in a solvent
inaccessible environment. The substrate binding site is formed
by a flat cavity with a volume of 420 Å3 (Fig. 3c). This cavity is
lined by a number of aromatic and aliphatic amino acids
(Fig. 4a,b), providing the highly hydrophobic environment pre-
dicted by substrate specificity and quantitative structure-activity
relationship (QSAR) studies17.
Adjacent to the substrate cavity is a separate, smaller
hydrophobic cavity (volume of 290 Å3) lined by residues
Phe 103, Pro 104, Trp 119, Leu 164, Leu 167, Phe 168, Leu 171,
Ile 199, Ile 316 and Tyr 326. This second cavity is situated
between the active site and the protein surface, and is shielded
from solvent by loop 99–112 (Fig. 3c). Residues Tyr 326, Ile 199,
Leu 171 and Phe 168 are the side chains that separate the two
cavities. These observations suggest a mechanism for admission
of the substrate into the active site that initially involves the
movement of loop 99–112 to open access to the smaller cavity
(termed the ‘entrance cavity’). After substrate reaches the
‘entrance cavity’, a transient movement of the four residues sepa-
rating the entrance from the substrate cavities must occur to
allow its diffusion into the active site. The total distance of sub-
strate migration from the surface of the entrance cavity to the
flavin ring is ∼20 Å. Loop 99–112 may function as a ‘gating
switch’ to the entrance cavity. The loop is situated proximal to
the membrane binding region (Fig. 3c), suggesting that sub-
strate must access the catalytic site from the protein surface ori-
ented towards the membrane.
Structural basis of catalysis
To analyze the mechanistic implications of the MAO B three-
dimensional structure, we have attempted to model the binding
of substrate in the active site. Benzylamine (Fig. 1b) was chosen
because there are considerable data on the interaction of this
substrate and its analogs with the enzyme17. Comparison of
flavoenzyme three-dimensional structures9 has revealed that the
substrate carbon atom undergoing flavin-dependent oxidation
binds in a highly conserved position in front of the flavin

24 nature structural biology • volume 9 number 1 • january 2002

 letters

Table 1 Crystallographic statistics flavin-dependent amine oxidases:

polyamine oxidase12 and trimethylamine
Data collection and phasing
dehydrogenase19. The catalytic signifi-
Data set Native Native PCMB
cance of the aromatic rings with respect to
Space group P1 C222 C222
the flavin-dependent amine oxidation is a
Cell axes (Å) 108.4, 132.4, 154.8 138.8, 224.3, 87.2 138.3, 222.2, 85.8
subject for future mechanistic studies.
Cell angles (°) 90.1, 90.5, 114.0
The volume of the benzylamine sub-
Resolution range (Å) 40.0–3.1 40.0–3.0 15.0–3.0
strate is ∼160 Å3, which is smaller than the
Completeness (%)1 98.4 (97.8) 95.3 (94.6) 99.1 (99.7)
© 2002 Nature Publishing Group http://structbio.nature.com

volume of the active site cavity (420 Å3).

Number of unique reflections 140,447 24,034 26,650
The modeling experiment highlights that
Rmerge1,2 0.088 (0.200) 0.128 (0.338) 0.126 (0.573)
the portion of the cavity on the rear side
Average redundancy1 3.8 (3.7) 8.1 (6.6) 6.3 (4.5)
with respect to the flavin ring is left unoc-
Rderiv3 0.174
cupied by the substrate (Fig. 4c). This
RCullis4 (centric / acentric) 0.78 / 0.86
implies that the cavity may allow an aro-
Phasing power5 (centric / acentric) 0.73 / 1.06
matic ring to bind at many positions, far-
ther or closer to the flavin. This feature
Refinement
explains the broad substrate specificity of
Resolution range (Å) 40–3.0
MAO B, which is able to oxidize several
Reflections (working / test set) 23,052 / 982
aromatic amines with side chains of dif-
Number of protein atoms6
ferent length (Fig. 1a).
chain A 3,959
As a step in the substrate oxidation reac-
chain B 3,932
tion, active site basic amino acid residues
Number of ligand atoms
have been suggested to function in H+
FAD 2 × 53
abstraction from the α-carbon atom of the
Pargyline 2 × 12
substrate –CH2-NH2 unit (Fig. 1b,c).
Rcryst7 0.230
Therefore, identifying side chains poten-
Rfree7 0.271
tially acting as active site bases is impor-
R.m.s. deviations
tant. Inspection of the substrate binding
Bond lengths (Å) 0.016
site shows that the aromatic cage residues
Bond angles (°) 2.3
Tyr 398 and Tyr 435 are the only side chains
NCS8 (Å) 0.09
that could potentially function in acid-base
Ramachandran plot9 (%)
catalysis. Other substrate binding residues
Allowed region 81
with an acidic or basic group are either not
Additionally allowed 18
properly positioned with respect to the
Generously allowed 0
Disallowed region 1
substrate (Tyr 60 and Tyr 188) or not con-
served in the sequence (Cys 172 and
1Values in parentheses are for reflections in the highest resolution shell. Tyr 326) (Fig. 4a,b). The possible partici-
2Rmerge = Σ|Ii – <I>| / ΣIi, where Ii is the intensity of ith observation and <I> is the mean intensity of pation of Tyr 398 and Tyr 435 as active site
the reflection.
3R
deriv = Σ||FPH| – |FP|| / Σ|FP|, where FP and FPH are the native and derivative structure factors, respec-
bases appears unlikely because recent
tively. mutagenesis data on the homologous
Cullis = Σ|||FPH| ± |FP|| – |FH|| / Σ||FPH| ± |FP||, where FP and FPH are defined as above and FH is the calcu-
4R
residues in human MAO A (Tyr 407 and
lated heavy atom structure factor.
5Phasing power = r.m.s. F / r.m.s. lack of closure (summed over all reflections used in the heavy
H
Tyr 444) show that the replacement of
atom refinement). either of these residues with Phe does not
6Out of a total of 520 amino acids for each MAO B monomer, chain A includes residues 4–500,
abolish catalytic activity20 but alters the
whereas chain B includes residues 4–496.
7R-factor = Σ|F – F | / Σ|F |, where F and F are the observed and calculated structure factor ampli- substrate specificity. These observations
o c o o c
tudes, respectively. Rcryst and Rfree were calculated using the working and test set, respectively. rule out the possibility that substrate oxi-
8R.m.s. deviation between all atoms of the two noncrystallographically symmetry-related
dation involves proton abstraction from
monomers present in the asymmetric unit of the C222 crystal form. Tight NCS restraints were
applied throughout the refinement.
the substrate by an active site amino acid
9Analyzed using PROCHECK35. The residues (φ,ψ values in subunit A and B) laying in the disallowed residue. This is consistent with the mecha-
regions of the Ramachandran plot are: Lys 52 (77°, –70° and 70°, –69°), Ala 346 (38°, –131° and 44°, nism of MAO catalysis recently proposed
–139°), Tyr 398 (50°, –87° and 50°, –93°) and Asp 419 (53°, –92° and 55°, –94°). from QSAR studies on MAO A18.

N5–C4a locus9. On this basis, the benzylamine methylene car-
bon was positioned 3.6 Å from flavin N5. The orientation of the
aromatic ring was restricted by the flat shape of the cavity
(Fig. 4c). As a result, the amine binds between the phenolic side
chains of Tyr 398 and Tyr 435 (edge to edge distance of 7.8 Å).
These residues, together with the flavin, form an aromatic caged
environment (Fig. 4a,c) that is responsible for recognition of the
amino group. No interaction of the substrate nitrogen atom with
any anionic residues is detected, which agrees with the known
preference of MAO to bind the deprotonated substrate18. An aro-
matic cage similar to that of MAO B is also observed in two other
Substrate specificities of MAOs
Human MAO A and MAO B share a high sequence identity
(∼70%) but differ in their substrate specificities. MAO B mainly
acts on small exogenous amines, whereas MAO A carries out the
degradation of bulkier endogenous amine neurotransmitters,
such as serotonin (Fig. 1a). The differences between human
MAO B and MAO A in residues lining the substrate cavity
(Fig. 4b) include Leu 171 (MAO B)/Ile 180 (MAO A),
Cys 172/Asn 181, Ile 199/Phe 208 and Tyr 326/Ile 335. These
amino acids are located in the rear of the substrate cavity
(Fig. 4a,b) in van der Waals contact with the benzene ring of

nature structural biology • volume 9 number 1 • january 2002 25

 letters
pargyline. Three of the four residues (Leu 171, Ile 199 and
Tyr 326) separate the substrate cavity from the entrance cavity.
Thus, side chain changes in these positions affect not only steric
accomodation of the substrate but may also lead to alterations in
the separation of the substrate cavity from the entrance cavity.
By altering the side chains of these residues, the two cavities may
fuse to form a single larger cavity that would accommodate larg-
er substrates and inhibitors, as observed for MAO A. Mutations
© 2002 Nature Publishing Group http://structbio.nature.com
of the cavity-separating residues in MAO B alter the substrate
and inhibitor specificities to resemble those of MAO A21,22. Taken
together, these data suggest that variations in substrate and
inhibitor specificities between human MAO A and MAO B
mainly result from the fine tuning of the size and shape of the
hydrophobic substrate cavity and its steric relation with the
entrance cavity.
These structural insights into human MAO B should be valu-
able in future inhibitor design that would specifically target each
form of MAO. The demonstration of an entrance cavity that con-
nects the surface of the protein to the substrate cavity in MAO B
may prove an additional useful site for the future design of
reversible inhibitors. This site may account for the divergent
QSAR results reported with different classes of MAO B inhibitors.
Methods
Crystallization and data collection. Recombinant human liver
MAO B was expressed and purified as described13. Two crystal forms
were obtained by employing different detergents (Table 1). Triclinic
crystals were grown from a solution containing 2 mg ml–1 MAO B in
25 mM potassium phosphate, pH 7.5, and 2.6 mM lauryldimethyl-
amine oxide (LDAO). Orthorhombic crystals (space group C222) were
obtained from a solution containing 2 mg ml–1 pargyline-inhibited
MAO B in 25 mM potassium phosphate, pH 7.5, and 8.5 mM n-dode-
cyl-N,N-dimethyl-3-amino-1-propanesulfonate (Zwittergent 3-12). In
both cases, crystals were grown by vapor diffusion methods at 4 °C
with a precipitant solution consisting of 12% (w/v) PEG 4000,
100 mM N-[2-acetamido]-2-iminodiacetic acid (ADA), pH 6.5, and
70 mM lithium sulphate.
Diffraction data were collected at 100 K at beamlines ID14-EH2
and ID14-EH3 of ESRF (Grenoble, France) and B7WB of DESY/EMBL
(Hamburg, Germany). Data were integrated with MOSFLM23 and
scaled using the CCP4 suite24 (Table 1). Crystals that grew with the
LDAO detergent belong to space group P1. Attempts to process the
data in higher symmetry space groups failed.
Structure solution and refinement. A heavy-atom derivative
was obtained by soaking a C222 crystal in a p-chloromercuribenzoic
acid (PCMB)-saturated solution. Six heavy-atom sites were found
using SHELXS-97 (ref. 25) and refined with the program MLPHARE24
(Table 1). The resulting electron density map was used to identify
the position and orientation of the noncrystallographic two-fold
axis using GLRF26, allowing phase refinement by two-fold averaging
and solvent flattening. The two-fold averaged map was employed
in molecular replacement calculations using AMoRE27 to find the
transformations that relate the dimer of the orthorhombic form to
the protein molecules present in the unit cell of triclinic crystals. The
10 solutions indicate that five MAO B dimers are present in the P1
unit cell. On this basis, multicrystal 12-fold averaging was per-
formed with DMMULTI28, producing a map of excellent quality
(Fig. 2) that easily allowed the building of the initial model by the
program O29. The atomic coordinates were subjected to REFMAC
maximum likelihood refinement30 using the data for the C222 crys-
tal form. Given the high quality of the experimental electron densi-
ty map, the multicrystal 12-fold averaged phases were included as
additional restraints. Moreover, the two noncrystallographic sym-
26
metry (NCS)-related subunits were subjected to tight NCS restraints
throughout the refinement. The final model (residues 4–500 for
monomer A and 4–496 for monomer B) has an R-factor of 23.0%
and an Rfree of 27.1%, with good stereochemical parameters
(Table 1). Four residues in each subunit are in disallowed regions of
the Ramachandran plot, one of which is Tyr 398 of the active center
(Table 1). Refinement of the model in the triclinic crystal form is cur-
rently in progress and will be described elsewhere together with a
more detailed analysis of MAO B structure. Volume calculations
were performed with VOIDOO31 using a probe radius of 1.4 Å.
Drawings were produced using MOLSCRIPT32, BOBSCRIPT33 and
DINO34.
Coordinates. Atomic coordinates have been deposited in the
Protein Data Bank (accession number 1GOS).
Acknowledgments
This work was supported by grants from the National Institute of General
Medical Sciences of the NIH, the Consiglio Nazionale delle Ricerche and Agenzia
Spaziale Italiana. We thank the staff of the EMBL/DESY and ESRF facilities for
help during the data collection. The European Union provided support through
the Human Capital Mobility Program to Large Scale Installations Project. We
thank A. Coda, B. Curti, M. Rizzi and R. van den Heuvel for helpful discussions.
Correspondence should be addressed to A.M. email: mattevi@ipvgen.unipv.it or
D.E.E. email: dedmond@bimcore.emory.edu
Received 17 August, 2001; accepted 29 October, 2001.
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