Vous êtes sur la page 1sur 7


DOI: 10.1002/chem.200901645

The Interaction of Zinc(II) and Hydroxamic Acids and a Metal-Triggered

Lossen Rearrangement

Lucie Duchčkov and Jana Roithov*[a]

Dedicated to Professor Jan Schraml on the occasion of his 70th birthday

Abstract: The structure and reactivity acetic acid is present in its neutral vide experimental evidence for the pro-
of a complex of zinc(II), water, acetic form. The binding energies of the li- posed mode of operation of drugs
acid, and acetohydroxamic acid, in gands towards zinc increase in the fol- based on hydroxamic acids. Further-
which one of the acids is deprotonated, lowing order: water < acetic acid < ace- more, coordinatively unsaturated com-
is investigated by means of mass spec- tohydroxamic acid. The structure of plexes of zinc and acetohydroxamic
trometry, labeling studies, and density the complex and its fragmentation pro- acid undergo a zinc-assisted Lossen re-
functional calculations to unravel the arrangement followed by elimination
exceptional binding properties of hy- of water if acetohydroxamic acid is
Keywords: density functional calcu-
droxamic acids towards zinc-containing present as a neutral ligand, or by loss
lations · hydroxamic acid · Lossen
enzymes at the molecular level. It is of methylisocyanate if acetohydroxa-
rearrangement · mass spectrometry ·
shown that acetohydroxamic acid is de- mic acid is deprotonated.
protonated in the complex, whereas

Introduction The structures of the active sites of many zinc-containing

enzymes show similar patterns consisting of a zinc cation co-
Zinc is one of the most frequently occurring metals in ordinated by glutamate and several histidine residues.[13] For
metalloenzymes. For example, zinc proteinases represent a example, the active sites of thermolysin or carboxypeptida-
broad family of enzymes participating in the biosynthesis se A are composed of a zinc(II) cation coordinated by two
and metabolism of proteins. A malfunction of proteinases histidines, glutamate, and a molecule of water, which partici-
can lead to development of serious diseases such as arthritis, pates in the hydrolysis of peptide bonds.[14, 15]
hypertension, or cancer.[1–3] Current research in this area is More detailed structural experiments using X-ray crystal-
concentrated towards the development of suitable inhibitors lography performed on several zinc-containing enzymes re-
of matrix metalloproteinases and histone deacetylases.[4–6] vealed that the coordination mode of glutamate can differ in
Both groups of enzymes play significant roles in the growth that it can be bound either as a monodentate or a bidentate
and spread of tumors.[1] The development of new drugs is in- ligand. This dichotomy has been explained by extensive the-
spired by the investigation of enzyme-inhibitor complexes, oretical studies by Ryde,[16] who showed that the coordina-
which can moreover bring valuable information about the tion mode of a carboxylic acid towards zinc strongly de-
activity of enzymes and the structures of the reactive com- pends on the other ligands bound to the metal. Not unex-
plexes.[7–12] pectedly, the more coordinatively saturated the zinc center
is, the more the bidentate binding mode is energetically dis-
favored. Moreover, a second solvation shell through hydro-
[a] L. Duchčkov, Dr. J. Roithov gen bonding towards the carboxylate group can also contrib-
Faculty of Science, Department of Organic and Nuclear Chemistry ute to the destabilization of the bidentate bonding. The
Charles University in Prague, Hlavova 2030/8 energy differences between both binding modes are, howev-
128 43 Prague 2 (Czech Republic)
Fax: (+ 420) 221951326
er, very small and therefore it is expected that dynamic
E-mail: roithova@natur.cuni.cz changes in bonding can contribute to the reactivity of
Supporting information for this article is available on the WWW zinc(II) centers in enzymes. For example, in a positively
under http://dx.doi.org/10.1002/chem.200901645. charged model of an active site composed of the zinc dicat-

Chem. Eur. J. 2009, 15, 13399 – 13405  2009 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim 13399
ion, two histidine molecules, acetate, and water, the mono-
dentate and bidentate modes of binding of carboxylates to-
wards zinc differ only by 10 kJ mol 1.
The mode of action of inhibitors for the most carefully
studied zinc proteinases, thermolysin and carboxypeptida-
se A, consists of a replacement of the water molecule ligated
at the zinc center by the inhibitor. Among the most potent
inhibitors are hydroxamic acids,[4, 17, 18] which are usually
more efficient inhibitors of zinc-containing enzymes than
their analogues based on carboxylates, phosphinates, or
others. It has been proposed that one of the unique features
of hydroxamic acids is their low acidity (in contrast to car-
boxylic or phosphinic acids). In fact, with typical pKa values
around 8.5, they are not deprotonated at physiological pH,
which means that they approach the active sites as neutral
molecules. Instead, deprotonation to form an effective che-
lating[19–21] hydroxamate group takes places only after coor-
dination to the zinc cation.[22, 23]

Results and Discussion

As outlined in the Introduction, the inhibition function of

hydroxamic acids on peptidases consists of the displacement
of the water molecule in the active center. We have modeled
the situation on the investigation of equilibria in a simple
gas-phase complex of zinc(II) with water, acetic acid, and
acetohydroxamic acid, in which one of the acids is deproton-
will use the first formula in the following. The complex of a collision energy of 2.8 eV. The pressure of xenon in the collision cell
interest was generated from an aqueous solution of ZnSO4, was 1  10 4 mbar. The inset shows the m/z 130–190 range on an enlarged
acetic acid, and acetohydroxamic acid (Figure S2 in the Sup- scale. b) CID of the perdeuterated complex [ZnACHTUNGRE(CH3CONDO)-
porting Information). The collision-induced dissociation ACHTUNGRE(CD3COOD)ACHTUNGRE(D2O)] + at a collision energy of 6.5 eV. The pressure in the
collision cell was deliberately increased to promote consecutive fragmen-
(CID) of the parent complex shows competitive losses of a tations. c) CID of the [ZnACHTUNGRE(CH3CONHO)ACHTUNGRE(CD3COOH)ACHTUNGRE(H2O)] + complex
water molecule and neutral acetic acid (Figure 1a), in which (m/z 219) containing acetic acid with a perdeuterated methyl group at a
the first one is more abundant and appears at lower collision collision energy of 2.8 eV. The pressure of xenon in the collision cell was
energies (energy-dependent CID spectra are given in Fig- 1  10 4 mbar. The inset shows the m/z 130–190 range on an enlarged
ure S3 in the Supporting Information). At a higher collision
energy, consecutive losses of water and acetic acid can be
observed. In addition, we observe two minor channels, of
which the first one corresponds to the consecutive elimina- Additional experiments with the complex [Zn-
tion of two molecules of water and the second channel rep- ACHTUNGRE(CH3CONHO)ACHTUNGRE(CD3COOH)ACHTUNGRE(H2O)] + generated from a solu-
resents the loss of hydroxamic acid. tion of ZnSO4, CD3COOD, and CH3CONHOH in excess
While the consecutive losses of water and acetic acid lead H2O reveals interesting findings (Figure 1c). The first loss of
directly to the complex of zinc(II) and deprotonated aceto- water corresponds to unlabeled H2O, as expected. The
hydroxamic acid ([ZnACHTUNGRE(CH3CONHO)] + ), the loss of two second loss of water shows almost equal abundances of
molecules of water is more ambiguous. To confirm the frag- losses of H2O and HDO, which suggests that the elimination
mentation pattern, the same experiment has also been per- can occur from both acetohydroxamic acid (H2O) and acetic
formed with complex [ZnACHTUNGRE(CH3CONDO)ACHTUNGRE(CD3COOD)- acid (HDO). Significant H/D exchange is observed also for
ACHTUNGRE(D2O)] + generated from a solution of ZnSO4, CD3COOD, the loss of CD3COOH, which is accompanied by the loss of
and CH3CONHOH in excess D2O (note that both acidic hy- [C2H2D2O2]. As the loss of the first molecule of water does
drogen atoms of acetohydroxamic acid were exchanged by not show any H/D scrambling, the scrambling does not
deuterons). The fragmentation observed (Figure 1b) is fully occur in the parent complex, but only after the loss of the
in agreement with the results of the unlabeled experiment. water ligand. The loss of [C2H2D2O2] is therefore assigned
Thus, the second molecule of water is lost either from acetic to a combined loss of H2O and ketene CD2CO as suggested
or acetohydroxamic acid. in Scheme 1.

13400 www.chemeurj.org  2009 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Chem. Eur. J. 2009, 15, 13399 – 13405
Zinc(II) and Hydroxamic Acids
ination of intact acetohydroxa-
mic acid is negligible and only
traces can be observed at colli-
sion energies above 4 eV.
In summary, the CID experi-
ments clearly show that aceto-
hydroxamic acid is the most
strongly bound ligand in the
complex and that most proba-
Scheme 1. Proposed fragmentation pathway for the consecutive eliminations of H2O/HDO and H2O/CD2CO
+ bly it is present as a counterion.
Hence, the deprotonation
within the complex involves
An energy-dependent CID study of the labeled complex acetohydroxamic acid and not acetic acid, as would be ex-
[ZnACHTUNGRE(CH3CONHO)ACHTUNGRE(CD3COOH)ACHTUNGRE(H2O)] + (Figure 2) lends pected based on the pKa values of acetohydroxamic acid
strong support to the suggested mechanism. The first mole- (8.7) and acetic acid (4.8). The water molecule is the weak-
cule of water is lost already at nominally zero collision est bound ligand, followed by acetic acid. After the loss of
water, the direct elimination of acetic acid is disfavored as it
leads to a zinc complex with only one ligand ([Zn-
ACHTUNGRE(CH3CONHO)] + ) and a combined loss of water and aceto-
hydroxamic acid is not observed at all. Instead, acetohy-
droxamic or acetic acid can undergo rearrangements and
eliminate another molecule of water. Due to the necessary
rearrangements, these channels are kinetically disfavored
and the direct elimination of acetic acid prevails at larger
collision energies.
Possible structural isomers of the investigated complex
were explored by DFT calculations (Figure 3), in which we
considered possible isomers with deprotonated acetohy-
droxamic acid, acetic acid, and water. In agreement with the
deduction derived from our experiments, the deprotonation
of the hydroxamic acid leads to the most stable structures of
the complex (1 + –3 + ). The deprotonation occurs at the
oxygen atom, as is already known for the complexes of hy-
Figure 2. CID of [66ZnACHTUNGRE(CH3CONHO)ACHTUNGRE(CD3COOH)ACHTUNGRE(H2O)] + (m/z 221) droxamic acids and metals,[25] and not at the nitrogen atom,
and its dependence on collision energy. The sum of the abundances of as in the isolated hydroxamate anion in the gas phase.[26]
the parent ion and all fragments is set to 1. The pressure of xenon in the
The most stable isomer 1 + bears neutral acetic acid as a
collision cell was 1  10 4 mbar. Elimination of H2O (m/z 203) is in black,
and that of CD3COOH (m/z 158, amplified by 5) in brown. Sequential monodentate ligand in the syn configuration (Zn-O-C-O di-
eliminations of H2O and H2O (m/z 185, amplified by 10) are depicted in hedral angle  08) and the structure is additionally stabilized
cyan, H2O and HDO (m/z 184, amplified by 10) in blue, H2O and by a hydrogen bond between acetic acid and hydroxamate.
CD2CO (m/z 159, amplified by 5) in red, H2O and CH3COOH (m/z 140, The hydrogen bond brings stabilization of about 0.3 eV as
amplified by 5) in dark yellow, and finally, the trace elimination of
CH3CONHOH (m/z 146, amplified by 10) is depicted in green.
can be deduced from the comparison of relative energies of
isomers 1 + and 3 + . Without consideration of the hydrogen
bond, the anti configuration of acetic acid is slightly favored
energy (signal at m/z 203, appearance energy (AE) over the syn configuration (compare 2 + and 3 + , respective-
 0.2 eV). As a second channel, we observe elimination of ly), which is opposite to what was found for deprotonated
H2O from acetohydroxamic acid (signal at m/z 185, AE acetic acid.[16] Deprotonation at acetic acid leads to higher
 0.9 eV). A competition between the elimination of water lying isomers 4 + and 5 + and in the most stable configura-
and an elimination of intact acetic acid ( CD3COOH, m/z tions found, acetate acts as a bidentate ligand. Deprotona-
158) can be observed above AE  2.0 eV. Next, two coupled tion of water leads to energy-disfavored isomers 6 + and 7 + ,
fragmentations arise from initial elimination of H2O fol- which lie considerably higher in energy although they are
lowed by 1) rearrangement of the deuterated acetic acid stabilized by a network of hydrogen bonds.
ligand to HDO and CD2CO, and 2) elimination of either Hence, the experimental and theoretical results clearly
HDO or CD2CO (signals at m/z 184 and 159 with the AE demonstrate that the assumptions about the inhibition func-
values of 2.1 and 2.6 eV, respectively). At even larger colli- tion of hydroxamic acid are valid. Firstly, the coordination
sion energies, the rearrangement of acetic acid is overridden of the hydroxamic acid to the zinc acetate complex is fol-
by the consecutive elimination of H2O and a whole mole- lowed by a hydrogen migration from the hydroxamic acid to
cule of acetic acid (m/z 140, AE  3.6 eV). Notably, the elim- acetate. Secondly, hydroxamate is not only much more

Chem. Eur. J. 2009, 15, 13399 – 13405  2009 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim www.chemeurj.org 13401
J. Roithov and L. Duchčkov

The elimination of acetic acid

the complex of zinc acetohy-
droxamate with water ([Zn-
requires 1.14 eV. For compari-
son, the alternative complex
[ZnOH(CH3CONHOH)] + lies
0.56 eV higher in energy. Ace-
tohydroxamic acid is coordinat-
ed in both complexes, whether
deprotonated or neutral, as a
bidentate ligand. Finally, elimi-
nation of acetohydroxamic acid
from the parent complex re-
quires substantially higher
energy. Formation of [Zn-
ed with 1.99 eV endoergicity
and that of [ZnOH-
ACHTUNGRE(CH3COOH)] + with 2.29 eV.
Similarly to the results found
above, the acetic acid is coordi-
nated as a bidentate ligand if it
Figure 3. Isomers of [ZnACHTUNGRE(CH3CONHO)ACHTUNGRE(CH3COOH)ACHTUNGRE(H2O)] + optimized at the B3LYP/TZVP level of theory. is deprotonated, but as a mono-
The selected bond lengths are in . Zinc is in yellow, oxygen in red, nitrogen in green, and carbon and hydro- dentate ligand if it is in the neu-
gen atoms are in gray . tral form.

strongly bound to the zinc center than water, but also than Table 1. Energetics of possible fragmentations of the [ZnACHTUNGRE(CH3CONHO)-
the acetic acid. The elimination of the water molecule re- ACHTUNGRE(CH3COOH)ACHTUNGRE(H2O)] + complex.
quires 0.8 eV (Figure 4, Table 1). The resulting fragment ion Products DE0relK [eV][a] DG298
bears again preferentially deprotonated hydroxamic acid [ZnACHTUNGRE(CH3CONHO)ACHTUNGRE(CH3COOH)] + H2O
1.26 0.81
and neutral acetic acid, and the latter is coordinated as a [ZnACHTUNGRE(CH3COO)(CH3CONHOH)] + + H2O 1.39 0.97
monodentate ligand in the syn configuration. The alternative [ZnACHTUNGRE(CH3CONHO)ACHTUNGRE(H2O)] + + CH3COOH 1.63 1.14
[Zn(OH)(CH3CONHOH)] + + CH3COOH 2.20 1.70
isomer with acetate and neutral hydroxamic acid lies [ZnACHTUNGRE(CH3COO)ACHTUNGRE(H2O)] + + CH3CONHOH 2.53 1.99
0.15 eV higher in energy. [Zn(OH)CH3COOH)] + + CH3CONHOH 2.87 2.29
[a] Energies are given relative to E0relK([ZnACHTUNGRE(CH3CONHO)ACHTUNGRE(CH3COOH)-
ACHTUNGRE(H2O)] + ) = 2368.688398 hartree; E0relK is a sum of the total electronic
energy and zero-point vibrational energy of a given species. [b] Energies
are given relative to G298 rel ([ZnACHTUNGRE(CH3CONHO)ACHTUNGRE(CH3COOH)ACHTUNGRE(H2O)] ) =
K +

2368.732373 hartree; G298

is the Gibbs energy of a given species calcu-
lated at 298 K.

We note in passing that the calculated binding energies do

not fully correlate with the experimentally determined ap-
pearance energies. The appearance energy for the elimina-
tion of water (0.2 eV) is lower than the theoretical threshold
(0.81 eV), which is most probably due to the internal energy
deposited in the parent complex upon its generation and the
kinetic energy distribution of the parent-ion beam. On the
other hand, the remaining fragmentations are affected by
the kinetic factors and therefore the appearance energies
Figure 4. Possible fragmentation of the [ZnACHTUNGRE(CH3CONHO)ACHTUNGRE(CH3COOH)-
are substantially higher than the theoretical values. In the
ACHTUNGRE(H2O)] + complex. Zinc is in yellow, oxygen in red, nitrogen in green, and further discussion, we therefore consider the theoretical
carbon and hydrogen atoms are in gray values, which more precisely reflect the actual binding ener-

13402 www.chemeurj.org  2009 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Chem. Eur. J. 2009, 15, 13399 – 13405
Zinc(II) and Hydroxamic Acids
gies and are not compromised by the kinetics of fragmenta- at higher collision energies (Figure 5b). Note that analogous
tion. Lossen rearrangement also accounts for the second water
To further substantiate the interaction between zinc(II) molecule loss from the [ZnACHTUNGRE(CH3CONHO)ACHTUNGRE(CH3COOH)-
and hydroxamic acids, we have investigated the reactivity ACHTUNGRE(H2O)] + complex (besides that which is due to the rear-
within the complex of zinc(II), acetohydroxamic acid, and rangement of acetic acid; see above).
acetohydroxamate (Figure 5). The collisional activation of The same sequence of rearrangements can be also pro-
posed for the deprotonated acetohydroxamic acid
(Scheme 2, pathway b).[28] In this case, only elimination of
methylisocyanate is expected. To this end, the fragmentation
of the acetohydroxamate ion bound to the zinc cation was
probed in a CID experiment of the [ZnACHTUNGRE(CH3CONHO)-
ACHTUNGRE(phen)] + complex (phen = phenanthroline), in which the
phenanthroline ligand preserves a favorable coordination
environment for zinc(II) but cannot deliver a proton. The
spectrum shows the elimination of CH3NCO (methylisocya-
nate or the corresponding nitrene) as a dominant fragmenta-
tion pathway, which is fully consistent with the formation of
[Zn(OH)ACHTUNGRE(phen)] + (Figures S5–S7 in the Supporting Infor-
mation). Elimination of lighter fragments is not observed.
Another fragmentation of [Zn-
ACHTUNGRE(CH3CONHO)(CH3CONHOH)] + observed at higher colli-
sion energies corresponds to a loss of ketene (CH2CO). A
probable mechanistic pathway involves a hydrogen rear-
rangement from the methyl group to the nitrogen atom
Figure 5. CID of the [ZnACHTUNGRE(CH3CONHO)(CH3CONHOH)] + complex (m/z within the neutral ligand (Scheme 2, pathway c), which leads
213) at the center-of-mass collision energy of a) 1.0 and b) 8.6 eV. to a complex containing ketene and hydroxylamine as neu-
tral ligands.
Recently, a new synthetic approach using a base-catalyzed
the complex leads first to the elimination of a water mole- Lossen rearrangement has been reported.[29] It has been
cule (Figure 5a) and, only at higher collision energies, a loss shown that aromatic hydroxamic acids in basic nonaqueous
of an intact molecule of acetohydroxamic acid can compete solutions rearrange to yield aromatic amines. The basicity is
efficiently (Figure 5b). assured by the addition of K2CO3 or similar salts. Earlier it
As to the mechanism, the loss of water from a molecule has already been shown that acidic salts of hydroxamic acids
of hydroxamic acid corresponds formally to the formation and alkali metals also undergo Lossen rearrangement under
of a nitrene in the first step, which undergoes a rearrange- mild conditions.[25] Our results suggest that the rearrange-
ment to yield isocyanate. Thus, presumably in the neutral ment of acetohydroxamic acid (deprotonated or neutral) co-
ligand of the [ZnACHTUNGRE(CH3CONHO)(CH3CONHOH)] + com- ordinated to the metal ions is subject to only a very small
plex, a hydrogen rearrangement from the nitrogen atom to energy barrier. Consequently, it can be proposed that in the
the hydroxyl group leads to the
formation of a complex in
which zinc bears two neutral li-
gands, water and the acylni-
trene CH3CON (Scheme 2,
pathway a). The latter can un-
dergo a rearrangement to yield
a more stable methylisocyanate
(CH3NCO). In analogy to the
organic variant of this reac-
tion,[27] this reaction can be re-
ferred to as a zinc-assisted
Lossen rearrangement of aceto-
hydroxamic acid in the gas
phase. The complex formed in
this way can lose a water ligand
as observed in the spectrum,
but it can also lose methyliso-
cyanate, which can be observed Scheme 2. Possible rationale for a zinc-assisted Lossen rearrangement.

Chem. Eur. J. 2009, 15, 13399 – 13405  2009 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim www.chemeurj.org 13403
J. Roithov and L. Duchčkov

rearranged complex of zinc, methylisocyanate, and water, manifold during the experiments was in the order of (3–10)  10 mbar
hydrolysis can occur to yield methylamine and CO2. Thus, it and that of xenon in the collision cell around (1–2)  10 4 mbar.

remains for further investigation to address this particular The collision-induced dissociation (CID) of the mass-selected ions can be
performed at variable collision energies adjusted by the potential differ-
reaction and to answer the challenging question of whether ence between Q1 and O in the range of 0–196 V, whereas the potential at
a general metal-triggered Lossen rearrangement is opera- O2 was set to the sum of the offsets of Q1 and O. The kinetic energy res-
tive. In the broader context of hydroxamic acids as biologi- olution of the mass-selected monocations is typically (1.1  0.1) eV (full-
cally active compounds, the facile Lossen rearrangement ap- width at half-maximum) and the inflex point on the rise of the kinetic
energy profile was taken as the nominal zero of the collision-energy scale
pears to be an interesting perspective. Such a metal-trig-
(Figure S1 in the Supporting Information). The collision energies (Elab)
gered rearrangement would lead to the formation of highly were converted to the center-of-mass frame energies (ECM = Elab  m/ACHTUNG-
reactive isocyanates, which are likely to interact with some TRENUG(m+M), in which m and M are masses of xenon and the parent ion, re-
of the neighboring groups. spectively). Variation of the collision energy leads to breakdown dia-
grams that enable qualitative determination of appearance energies
(AEs) of the various fragmentation channels.[31] Threshold behavior was
analyzed empirically by fitting a sigmoid function at the rise of an ion
Conclusion yield and the appearance energies were obtained as the linear extrapola-
tion of the tangent at the inflex point of the sigmoid function to the base-
The gas-phase experiments with a model complex of zinc(II) line. The AEs determined in this way are to be understood as a qualita-
tive lead for the discussion and for the comparison of individual fragmen-
with water, acetic acid, and acetohydroxamic acid (one of
tation channels and not as absolute binding energies of the corresponding
the acids being deprotonated) reveal that water is the weak- fragments in the complex.
est bound ligand followed by acetic acid, and the strongest The ionic products formed in the octopole were mass-analyzed by scan-
bound ligand is acetohydroxamic acid. Accordingly, acetohy- ning Q2. The mass resolutions of both (Q1 and Q2) were adjusted to
droxamic acid is in the deprotonated form, which is also fully resolve the nominal masses of all ionic species studied. Ion abun-
confirmed by the theoretical calculations. This finding sup- dances were determined by using a Daly-type detector operated in the
counting mode. The experiments in the manifold were performed at
ports the suggested mode of operation of drugs based on hy- 40 8C. Usually, several hundreds of scans were accumulated to improve
droxamic acids, which are supposed to approach the zinc- the signal-to-noise ratio.
containing active centers of enzymes, displace a water The calculations were performed using the density functional method
ligand, and upon coordination, transfer the acid proton to a B3LYP[32–35] in conjunction with the TZVP basis sets[36] as implemented
nearby glutamate. in the Gaussian 03 suite.[37] For all optimized structures, frequency analy-
ses at the same level of theory were used to assign them as genuine
In addition, we have investigated fragmentation patterns
minima or transition structures on the potential-energy surface (PES) as
of zinc complexes with hydroxamic acid being bonded as a well as to calculate zero-point vibrational energies (ZPVEs). The relative
neutral or anionic ligand. The major fragmentation path- energies (Erel) of the structures given here refer to energies at 0 K. The
ways correspond in both cases to a zinc-assisted Lossen rear- optimized geometries are listed in the Supporting Information.
rangement of the hydroxamic acid followed by the elimina-
tion of water or isocyanate. The elimination of water is pre-
ferred for the complexes in which the hydroxamic acid acts
as a neutral ligand, whereas in the complexes with deproton-
ated hydroxamic acid an elimination of isocyanate is ob-
served leaving a hydroxyl bound to zinc as a counterion. The authors thank the Grant Agency of the Academy of Sciences of the
Czech Republic (KJB400550704), the Ministry of Education of the Czech
Republic (MSM0021620857, RP MSMT 14/63), and the Charles Universi-
ty Grant Agency (259029) for their support. J.R. thanks Spectronex s.r.o.
Experimental Section (Prague) for providing the TSQ 7000 instrument and M. Novk for its in-
The experiments were performed using a TSQ 7000 mass spectrometer,
which was equipped with electrospray ionization (ESI) and atmospheric
pressure chemical ionization (APCI) sources; only ESI is used in this [1] F. Grams, M. Crimmin, L. Hinnes, P. Huxley, M. Pieper, H. Tsche-
study. The analyzer has a quadrupole–octopole–quadrupole (QOQ) con- sche, W. Bode, Biochemistry 1995, 34, 14012.
figuration, which permits a variety of MS/MS experiments. The studied [2] M. A. Holbert, R. Marmorstein, Curr. Opin. Struct. Biol. 2005, 15,
ions were generated from an aqueous solution of zinc sulfate, acetic acid, 673.
and acetohydroxamic acid, if not mentioned otherwise. The generated [3] H. Y. Lin, C. S. Chen, S. P. Lin, J. R. Weng, C. S. Chen, Med. Res.
ions are led through a heated capillary (heated to 250 8C) and tube lens Rev. 2006, 26, 397.
towards a small guide (radio frequency (rf)-only) quadrupole. The adjust- [4] M. Dokmanovic, C. Clarke, P. S. Marks, Mol. Cancer Res. 2007, 5,
ment of voltages on the capillary and tube lens allows for the modifica- 981.
tion of the ionization to soft or hard conditions; the soft conditions pro- [5] S. Ropero, M. Esteller, Mol. Oncol. 2007, 1, 19.
vide larger ionic clusters, whereas the hard conditions lead to the genera- [6] M. Whittaker, C. D. Floyd, P. Brown, A. J. H. Gearing, Chem. Rev.
tion of smaller desolvated ions.[30] The first analyzer quadrupole was used 1999, 99, 2735.
for mass analysis of ions formed in the ion source as well as for the mass [7] D. G. Hangauer, A. F. Monzingo, B. W. Matthews, Biochemistry
selection of the ions submitted to the subsequent unimolecular or bimo- 1984, 23, 5730.
lecular reactivity experiments. The octopole served as a collision cell, [8] C. L. Yan, Z. L. Xiu, X. H. Li, S. M. Li, C. Hao, H. Teng, Proteins
therefore it was placed in a housing separating the higher pressure colli- Struct. Funct. Bioinf. 2008, 73, 134.
sion region from the rest so as to minimize the diffusion of a collision gas [9] D. W. Christianson, W. N. Lipscomb, Acc. Chem. Res. 1989, 22, 62,
(usually xenon) to the high-vacuum manifold. The typical pressure in the and references therein.

13404 www.chemeurj.org  2009 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Chem. Eur. J. 2009, 15, 13399 – 13405
Zinc(II) and Hydroxamic Acids
[10] H. Feinberg, H. M. Greenblatt, V. Behar, C. Gilon, S. Cohen, A. [26] M. Decouzon, O. Exner, J.-F. Gal, P. Ch. Maria, J. Org. Chem. 1990,
Bino, G. Shoham, Acta Crystallogr. Sect. D 1995, 51, 428, and refer- 55, 3980.
ences therein. [27] L. Bauer, O. Exner, Angew. Chem. 1974, 86, 419; Angew. Chem. Int.
[11] W. L. Mock, H. Cheng, Biochemistry 2000, 39, 13945. Ed. Engl. 1974, 13, 376.
[12] S. P. Gupta, Chem. Rev. 2007, 107, 3042. [28] G. W. Adams, J. H. Bowie, R. N. Hayes, J. Chem. Soc. Perkin Trans.
[13] W. Stocker, F. Grams, U. Baumann, P. Reinemer, F.-X. Gomis-Ruth, 2 1991, 689.
D. B. McKay, W. Bode, Protein Sci. 1995, 4, 823, and references [29] Y. Hoshino, M. Okuno, E. Kawamura, K. Honda, S. Inoue, Chem.
therein. Commun. 2009, 2281.
[14] B. W. Matthews, Acc. Chem. Res. 1988, 21, 333, and references [30] N. B. Cech, C. G. Enke, Mass Spectrom. Rev. 2001, 20, 362.
therein. [31] D. Schrçder, M. Engeser, M. Brçnstrup, C. Daniel, J. Spandl, H.
[15] M. W. Y. Szeto, J. I. Mujika, J. Zurek, A. J. Mulholland, J. N. Harvey, Hartl, Int. J. Mass Spectrom. 2003, 228, 743.
J. Mol. Struct. 2009, 917, 106. [32] S. H. Vosko, L. Wilk, M. Nusair, Can. J. Phys. 1980, 58, 1200.
[16] U. Ryde, Biophys. J. 1999, 77, 2777. [33] C. Lee, W. Yang, R. G. Parr, Phys. Rev. B 1988, 37, 785.
[17] C. J. Marmion, D. Griffith, K. B. Nolan, Eur. J. Inorg. Chem. 2004, [34] A. D. Becke, Phys. Rev. A 1988, 38, 3098.
3003. [35] B. Miehlich, A. Savin, H. Stoll, H. Preuss, Chem. Phys. Lett. 1989,
[18] D. Wang, P. Helquist, O. Wiest, J. Org. Chem. 2007, 72, 5446. 157, 200.
[19] J. R. Somoza, R. J. Skene, B. A. Katz, C. Mol, J. D. Ho, A. J. Jen- [36] A. Schfer, C. Huber, R. Ahlrichs, J. Chem. Phys. 1994, 100, 5829.
nings, C. Luong, A. Arvai, J. J. Buggy, E. Chi, J. Tang, B. C. Sang, E. [37] Gaussian , Revision C.02, M. J. Frisch, G. W. Trucks, H. B. Schlegel,
Verner, R. Wynands, E. M. Leahy, D. R. Dougan, G. Snell, M. G. E. Scuseria, M. A. Robb, J. R. Cheeseman, J. A. Montgomery, Jr.,
Navre, M. W. Knuth, R. V. Swanson, D. E. McRee, L. W. Tari, Struc- T. Vreven, K. N. Kudin, J. C. Burant, J. M. Millam, S. S. Iyengar, J.
ture 2004, 12, 1325. Tomasi, V. Barone, B. Mennucci, M. Cossi, G. Scalmani, N. Rega,
[20] A. Vannini, C. Volpari, G. Filocamo, E. C. Casavola, M. Brunetti, D. G. A. Petersson, H. Nakatsuji, M. Hada, M. Ehara, K. Toyota, R.
Renzoni, P. Chakravarty, C. Paolini, R. De Francesco, P. Gallinari, Fukuda, J. Hasegawa, M. Ishida, T. Nakajima, Y. Honda, O. Kitao,
C. Steinkuhler, S. Di Marco, Proc. Natl. Acad. Sci. USA 2004, 101, H. Nakai, M. Klene, X. Li, J. E. Knox, H. P. Hratchian, J. B. Cross,
15064. V. Bakken, C. Adamo, J. Jaramillo, R. Gomperts, R. E. Stratmann,
[21] D. P. Dowling, S. L. Gantt, S. G. Gattis, C. A. Fierke, D. W. Christi- O. Yazyev, A. J. Austin, R. Cammi, C. Pomelli, J. W. Ochterski, P. Y.
anson, Biochemistry 2008, 47, 13 554. Ayala, K. Morokuma, G. A. Voth, P. Salvador, J. J. Dannenberg,
[22] M. Izquierdo-Martin, R. L. Stein, J. Am. Chem. Soc. 1992, 114, 325. V. G. Zakrzewski, S. Dapprich, A. D. Daniels, M. C. Strain, O.
[23] J. B. Cross, J. S. Duca, J. J. Kaminski, V. S. Madison, J. Am. Chem. Farkas, D. K. Malick, A. D. Rabuck, K. Raghavachari, J. B. Fores-
Soc. 2002, 124, 11004. man, J. V. Ortiz, Q. Cui, A. G. Baboul, S. Clifford, J. Cioslowski,
[24] An isotopomer with 66Zn, [66ZnACHTUNGRE(CH3CONHO)ACHTUNGRE(CD3COOH)ACHTUNGRE(H2O)] + B. B. Stefanov, G. Liu, A. Liashenko, P. Piskorz, I. Komaromi, R. L.
(m/z 221), was used in the CID experiment due to an impurity in Martin, D. J. Fox, T. Keith, M. A. Al-Laham, C. Y. Peng, A. Na-
the source spectra with m/z 201 and its possible cluster with water nayakkara, M. Challacombe, P. M. W. Gill, B. Johnson, W. Chen,
(m/z 219), which might contribute to the CID of [64Zn- M. W. Wong, C. Gonzalez, J. A. Pople, Gaussian, Inc., Wallingford
ACHTUNGRE(CH3CONHO)ACHTUNGRE(CD3COOH)ACHTUNGRE(H2O)] + and affect the water-loss chan- CT, 2004.
[25] J. Podlaha, I. Csaøov, L. Soukupova, J. Schraml, O. Exner, J. Received: June 16, 2009
Chem. Soc. 1998, 520. Published online: November 20, 2009

Chem. Eur. J. 2009, 15, 13399 – 13405  2009 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim www.chemeurj.org 13405