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Chemistry of the crosslinking of collagen during tanning.

The materials commonly used for crosslinking collagen as part of the process of converting animal hides into leather fall into three main groups: mineral tannages, aldehyde tannages, and "vegetable" tannages. The most important mineral crosslinking agents are hydrated basic chromium III sulfate complexes. These compounds form extended polynuclear coordination complexes containing hydroxol, oxo, and sulfato bridges into which ionized carboxyl groups on collagen enter readily as coordinating ligands accomplishing crosslinking. On pH adjustment and partial drying, highly stable complexes are formed with oxo bridges predominating and protein amide groups entering the coordination complex. The aldehyde tannages proceed through aldehyde condensation reactions with collagen amino groups to give alpha-hydroxyamines which can condense with other collagen amine groups to effect crosslinking. The vegetable type tanning agents, whether natural plant extracts or synthesized, are complex, high molecular weight polyhydroxy compounds that do not rely on crosslinking as such to be effective. Their effectiveness appears to depend on other properties. This and additional information concerning these commerical tannages are reviewed.

Aldehydes↗

Exploring the use of the tripeptide Gly-Gly-his as a selective recognition element for the fabrication of electrochemical copper sensors.

The modification of electrodes with the tripeptide Gly-Gly-His for the detection of copper in water samples is described in detail. The tripeptide modified electrode was prepared by first self-assembling 3-mercaptopropionic acid (MPA) onto the gold electrode followed by covalent attachment of the tripeptide to the self-assembled monolayer using carbodiimide coupling. The electrodes were characterized using electrochemistry, a newly developed mass-spectrometry method and quantum mechanical calculations. The mass spectrometry confirmed the modification to proceed as expected with peptide bonds formed between the carboxylic acids of the MPA and the terminal amine of the peptide. Electrochemical measurements indicated that approximately half the MPA molecules in a SAM are modified with the peptide. The peptide modified electrodes exhibited high sensitivity to copper which is attributed to the stable 4N coordinate complex the peptide formed around the metal ion to give copper the preferred tetragonal coordination. The formation of a 4 coordinate complex was predicted using quantum mechanical calculation and confirmed using mass spectrometry. The adsorption of the copper to the peptide modified electrode was consistent with a Langmuir isotherm with a binding constant of (8.1 +/- 0.4) 10(10) M(-1) at 25 degrees C.

Journal Article↗

HTP-1 coordinates synaptonemal complex assembly with homolog alignment during meiosis in C. elegans.

During meiosis, the mechanisms responsible for homolog alignment, synapsis, and recombination are precisely coordinated to culminate in the formation of crossovers capable of directing accurate chromosome segregation. An outstanding question is how the cell ensures that the structural hallmark of meiosis, the synaptonemal complex (SC), forms only between aligned pairs of homologous chromosomes. In the present study, we find that two closely related members of the him-3 gene family in Caenorhabditis elegans function as regulators of synapsis. HTP-1 functionally couples homolog alignment to its stabilization by synapsis by preventing the association of SC components with unaligned and immature chromosome axes; in the absence of the protein, nonhomologous contacts between chromosomes are inappropriately stabilized, resulting in extensive nonhomologous synapsis and a drastic decline in chiasma formation. In the absence of both HTP-1 and HTP-2, synapsis is abrogated per se and the early association of SC components with chromosomes observed in htp-1 mutants does not occur, suggesting a function for the proteins in licensing SC assembly. Furthermore, our results suggest that early steps of recombination occur in a narrow window of opportunity in early prophase that ends with SC assembly, resulting in a mechanistic coupling of the two processes to promote crossing over.

Animals↗

The effect of kirromycin on the metal ion coordination in complexes of elongation factor Tu from Bacillus stearothermophilus as inferred from the 17O-55Mn superhyperfine interaction.

The binding of the antibiotic kirromycin to elongation factor Tu (EF-Tu) leads to a severe line broadening of the Q-band manganese EPR lines of EF-Tu.Mn, EF-Tu.Mn.GDP, as well as EF-Tu.Mn.GDP.Pi indicating that the coordination sphere of the metal ion is changed by this interaction. The number of coordinated water molecules was determined from the 17O-55Mn superhyperfine coupling observable in H2(17)O enriched water; it is pH-dependent in the EF-Tu.Mn.GDP complex, at pH 6.8 probably four water molecules are coordinated with the protein bound manganese, at pH 8.4 one of the water molecules is replaced by a functional group from the protein. Independent of the pH, probably four and five water molecules are bound to the metal ion in the EF-Tu.Mn and in the EF-Tu.Mn.GDP.Pi complex, respectively. Kirromycin does not influence the number of water molecules bound to EF-Tu.Mn.GDP and EF-Tu.Mn.GDP.Pi, but leads to an increase of the number of water molecules coordinated to the metal ion in EF-Tu.Mn. The 17O-55Mn superhyperfine coupling constant in Mn(H2O)6 was determined from the EPR-spectra as 0.24 mT.

Anti-Bacterial Agents↗

EPR study of ferric native prostaglandin H synthase and its ferrous NO derivative.

Purified prostaglandin H synthase (EC 1.14.99.1) apoprotein, a polypeptide of 72 kDA, was titrated with hemin and EPR spectra of high-spin ferric heme were observed at liquid-helium temperature. With up to one hemin per polypeptide, a signal at g = 6.6 and 5.4, rhombicity 7.5%, evolved owing to specifically bound, catalytic active heme. At higher heme/polypeptide ratios signals at g = 6.3 and 5.9 were observed which were assigned to non-specific heme with no catalytic function. In microsomes from ram seminal vesicles the native enzyme showed the signal at g = 6.7 and 5.2 which could not be increased by the addition of hemin. Cyanide, an inhibitor of the enzyme, reacted at lower concentrations with the specific heme abolishing its signal at g = 6.6 and 5.4. Higher concentrations of cyanide were needed for the disappearance of the signal of non-specific heme. The reduced enzyme reacted with NO and formed two types of NO complexes. A transient complex, with a rhombic signal at gx = 2.07, gz = 2.01 and gy = 1.97, was assigned to a six-coordinate complex. The final, stable complex showed an axial signal at g = 2.12 and g = 2.001 and was assigned to a five-coordinate complex, where the protein ligand was no longer bound to the heme iron. Neither type of signal showed a hyperfine splitting from nitrogen of histidine indicating the absence of a histidine-iron bond in the enzyme. From these results and the similarity of the EPR signal at g = 6.6 and 5.4 to the signal of native catalase (EC 1.11.1.6) we speculated that tyrosinate might be the endogenous ligand of the heme in prostaglandin H synthase.

Animals↗

A new approach in 1D and 2D 13C high-resolution solid-state NMR spectroscopy of paramagnetic organometallic complexes by very fast magic-angle spinning.

Novel 1D and multidimensional solid-state NMR (SSNMR) methods using very fast magic-angle spinning (VFMAS) (spinning speed > 20 kHz) for performing 13C high-resolution SSNMR of paramagnetic organometallic complexes are discussed. VFMAS removes a majority of 13C-1H and 1H-1H dipolar couplings, which are often difficult to remove by RF pulse techniques in paramagnetic complexes because of large paramagnetic shifts. In the first systematic approach using the unique feature of VFMAS for paramagnetic complexes, we demonstrate a means of obtaining well-resolved 1D and multidimensional 13C SSNMR spectra, sensitivity enhancements via cross polarization, and signal assignments, and applications of dipolar recoupling methods for nonlabeled paramagnetic organometallic complexes of moderate paramagnetic shifts ( approximately 800 ppm). Experimental results for powder samples of small nonlabeled coordination complexes at 1H frequencies of 400.2-400.3 MHz show that highly resolved 13C SSNMR spectra can be obtained under VFMAS, without requirements of 1H decoupling. Sensitivity enhancement in 13C SSNMR via cross polarization from 1H spins was demonstrated with an amplitude-sweep high-power CP sequence using strong RF fields ( approximately 100 kHz) available in the VFMAS probe. 13C CPMAS spectra of nonlabeled Cu(II)(dl-alanine)2.(H2O) and V(III)(acetylacetonate)3 (V(acac)3) show that it is possible to obtain high-resolution spectra for a small quantity ( approximately 15 mg) of nonlabeled paramagnetic organometal complexes within a few minutes under VFMAS. Experiments on Cu(II)(dl-alanine)2.(H2O) demonstrated that 1H-13C dipolar recoupling for paramagnetic organometal complexes can be performed under VFMAS by application of rotor-synchronous pi-pulses to 1H and 13C spins. The results also showed that signal assignments for 13CH, 13CH3, and 13CO groups in paramagnetic complexes are possible on the basis of the amount of 13C-1H dipolar dephasing induced by dipolar recoupling. Furthermore, the experimental 2D 13C/1H chemical-shift correlation NMR spectrum obtained for nonlabeled V(acac)3 exhibits well-resolved lines, which overlap in 1D 13C and 1H spectra. Signals for different chemical groups in the 2D spectrum are distinguished by the 13C-1H dipolar dephasing method combined with the 2D 13C/1H correlation NMR. The assignments offer information on the existence of nonequivalent ligands in the coordination complex in solids, without requiring a single-crystal sample.

Journal Article↗

An unusual eight-coordinated copper complex: bis(4,7-diphenyl-1,10-phenanthroline)dinitratocopper(II).

In the title monomer, [Cu(NO3)2(C24H16N2)2], the copper(II) cation is eight-coordinate within an octahedral-like polyhedron. The coordination polyhedron is formed by two chelating diphenylphenanthroline groups that define the highly distorted CuN4 equatorial plane and two weakly bound bidentate (chelating) nitrate groups in the apical or axial positions. The complex crystallizes in the monoclinic space group C2/c; a twofold axis passes through the copper(II) cation and bisects the two nitrate ligands. This gives the molecule C2 point-group symmetry, rendering only half of the molecule independent.

Journal Article↗

A dicoordinate palladium(0) complex with an unusual intramolecular eta(1)-arene coordination.

The reaction of (tmeda)PdMe2 with dcpBiph gives (dcpBiph)2Pd in high yield. The solid-state structure of (dcpBiph)2Pd reveals a bent P-Pd-P framework and an unusual eta1-arene interaction between the palladium and the distal ring of one of the biphenyl substituents. In solution, an additional conformer exists which does not show a pi-interaction with a biphenyl ring. The low-coordinate complex, (dcpBiph)2Pd, undergoes C-X oxidative addition reactions with PhX (X = I, Br, Cl). A minor product resulting from metalation of the biphenyl ring is also observed.

Journal Article↗

Bactericidal properties of hydrogen peroxide and copper or iron-containing complex ions in relation to leukocyte function.

Various combinations of hydrogen peroxide, reductant (ascorbic acid and superoxide ion), and copper or iron salts and their coordination complexes were examined to determine their cytotoxicity toward several bacteria with diverse metabolic capabilities and cell envelope structures. Four sets of bactericidal conditions were identified, comprising: (1) high concentration levels (5-100 mM) of H2O2 in the absence of exogenous metal ions and reductant; (2) ferrous or ferric coordination complexes plus enzymatically generated O2.- and H2O2 at relatively low steady-state concentration levels; (3) cupric ion plus low concentration levels of H2O2 (1 microM-1 mM) and ascorbate (10 microM-4 mM); (4) cuprous ion (or cupric ion plus ascorbate) in the absence of O2 and H2O2. Rates of losses in viabilities increased proportionately with increases in the concentration of H2O2 in metal-free environments and with each of the components in the Cu2+/ascorbate/H2O2 bactericidal assay system. Oxidant levels required for equivalent killing increased with increasing cell densities of the bacterial suspensions over the range investigated (2 x 10(7)-2 x 10(9) cfu/ml). Other experimental conditions or other combinations of reagents, most notably Fe3+/ascorbate/H2O2 systems, did not generate bactericidal environments. The patterns of response of the three organisms tested, Streptococcus lactis, Escherichia coli, and Pseudomonas aeruginosa, were similar, suggesting common bactericidal mechanisms. However, preliminary evidence suggests that the lethal lesions caused by the various bactericidal conditions are distinct: As discussed, each of the four bactericidal conditions could conceivably be attained within the phagosomes of leukocytes, although none has as yet been identified.

Anti-Bacterial Agents↗

Nonhaem complexes of FeIII stimulate cell attachment and growth by a mechanism different from that of serum, 2-oxocarboxylates, and haemproteins.

Most cell lines, even those producing their own growth factors, need a serum supplement when growing in several commonly used media. The requirement for serum to sustain attachment and growth in RPMI 1640 and MEM has been found to be met by a range of 2-oxocarboxylates, by diverse coordination complexes of FeIII, and by a variety of haem-containing proteins including catalase. The latter directly implicates H2O2 in the serum shift-down effects. H2O2 was found to accumulate in low serum media under normal laboratory lighting conditions to levels that were shown to be sufficient, when added to freshly prepared media, to explain the depressed cell performance. With the exception of some of the nonhaem FeIII coordination complexes, substances found to stimulate cell attachment and growth were capable of scavenging H2O2. This suggests that an important function of serum and the 2-oxocarboxylates (alpha-keto acids) frequently used as "nonessential" medium additives is to remove H2O2 produced photodynamically during the storage and manipulation of media containing a high content of riboflavin. However, the nonhaem FeIII complexes with saturated coordination shells, although capable of reducing photodynamic generation of H2O2 to a greater or lesser extent, have their prime effect by an unknown, intriguing mechanism, probably based on a common redox function.

Blood Physiological Phenomena↗

Structure of chlorobis(dicyclohexylphenylphosphine)gold(I).

[AuCl[C6H11)2(C6H5)P]2], Mr = 781.2, monoclinic, P2(1), a = 11.117 (4), b = 13.606 (5), c = 12.284 (4) A, beta = 109.59 (3) degrees, V = 1753 (2) A3, Z = 2, D chi = 1.48 g cm-3, lambda(Mo K alpha) = 0.71073 A, mu = 43.8 cm-1, F(000) = 792, T = 296 K, final R = 0.024 for 3239 unique observed reflections. The formation of complexes of the type LnAuX, where L is a phosphine, and X is halide or pseudohalide, shows considerable dependence on the nature of the phosphine. When L = Ph3P, three- and four-coordinated complexes are very easily prepared. However, for L = Cy3P, only linearly coordinated complexes could be isolated. The geometry of the title complex, with L = CyPhP, is best described as distorted trigonal planar. The P-Au-P angle is 158.24 (5) degrees while the P-Au-Cl angles are 92.63 (6) and 108.71 (6) degrees. Au-P distances are 2.300 (1) and 2.324 (1) A while Au-Cl is 2.744 (2) A.

Gold↗

Absolute configurational assignment of self-organizing asymmetric tripodal ligand-metal complexes.

The solution configuration of labile coordination complexes may be difficult to determine, even in cases in which the solid state structure is known. We have previously synthesized a series of chiral ligands which form pseudo-C3-symmetric complexes with ZnII and CuII salts that possess an available electrophilic coordination site. Molecular modeling of ZnII complexes of the chiral ligand N,N-bis[(2-quinolyl)methyl]-1-(2-pyridyl)ethanamine (alpha-MeBQPA) showed that the spatial arrangement of the heterocyclic arms is controlled by a substituent on one methylene arm, resulting in the adoption of an enantiomeric conformation displaying a propeller-like asymmetry. In this paper we report the application of the exciton chirality method to the determination of the conformation of asymmetric metal-ligand complexes in solution. There is a good correlation between the predicted and the observed Cotton effects, demonstrating that the geometry in solution closely resembles that predicted by computational simulations and those obtained by X-ray crystallographic studies of metal complexes with racemic and enantiomerically pure ligands. The X-ray crystallographic structure of the first optically pure complex in this series is reported.

Circular Dichroism↗

Automatic method for identifying reaction coordinates in complex systems.

To interpret simulations of a complex system to determine the physical mechanism of a dynamical process, it is necessary to identify the small number of coordinates that distinguish the stable states from the transition states. We develop an automatic method for identifying these degrees of freedom from a database of candidate physical variables. In the method neural networks are used to determine the functional dependence of the probability of committing to a stable state (committor) on a set of coordinates, and a genetic algorithm selects the combination of inputs that yields the best fit. The method enables us to obtain the first set of coordinates that is demonstrably sufficient to specify the transition state of the C(7eq)--> alpha(R) isomerization of the alanine dipeptide in the presence of explicit water molecules. It is revealed that the solute-solvent coupling can be described by a solvent-derived electrostatic torque around one of the main-chain bonds, and the collective, long-ranged nature of this interaction accounts for previous failures to characterize this reaction.

Alanine↗

Formation of ternary complexes by coordination of (diethylenetriamine)-platinum(II) to N1 or N7 of the adenine moiety of the antiviral nucleotide analogue 9.

The synthesis of (Dien)Pt(PMEA-N1), where Dien = diethylenetriamine and PMEA2- = dianion of 9-[2-(phosphonomethoxy)ethyl]adenine, is described. The acidity constants of the threefold protonated H3[(Dien)Pt(PMEA-N1)]3+ complex were determined and in part estimated (UV spectrophotometry and potentiometric pH titration): The release of the proton from the (N7)H+ site in H4[(Dien)Pt(PMEA-N1)]3+ occurs with a rather low pKa (= 0.52+/-0.10). The release of the proton from the -P(O)2(OH) group (pKa = 6.69+/-0.03) in H[(Dien)Pt(PMEA-N1)]+ is only slightly affected by the N1-coordinated (Dien)Pt2+ unit. Comparison with the acidic properties of the H[(Dien)Pt(PMEA-N7)]+ species provides evidence that in the (Dien)Pt(PMEA-N7) complex in aqueous solution an intramolecular, outer-sphere macrochelate is formed through hydrogen bonds between the -PO3(2-) residue of PMEA2- and a PtII-coordinated (Dien)NH2 group; its formation degree amounts to about 40%. The stability constants of the M[(Dien)Pt(PMEA-N1)]2+ complexes with M2+ = Mg2+, Ca2+, Ni2+, Cu2+ and Zn2+ were measured by potentiometric pH titrations in aqueous solution at 25 degrees C and I = 0.1 M (NaNO3). Application of previously determined straight-line plots of log K(M(R-PO3))M versus pK(H(R-PO3)H for simple phosph(on)ate ligands. R-PO3(2-), where R represents a non-inhibiting residue without an affinity for metal ions, proves that the primary binding site of (Dien)Pt(PMEA-N1) is the phosphonate group with all metal ions studied; in fact, Mg2+, Ca2+ and Ni2+ coordinate (within the error limits) only to this site. For the Cu[(Dien)Pt(PMEA-N1)]2+ and Zn[(Dien)Pt(PMEA-N1)]2- systems also the formation of five-membered chelates involving the ether oxygen of the -CH2-O-CH2-PO3(2-) residue could be detected; the formation degrees are about 60% and 30%, respectively. The metal-ion-binding properties of the isomeric (Dien)Pt(PMEA-N7) species studied previously differ in so far that the resulting M[(Dien)Pt(PMEA-N7)]2+ complexes are somewhat less stable, but again Cu2+ and Zn2+ also form with this ligand comparable amounts of the mentioned five-membered chelates. In contrast, both M[(Dien)Pt(PMEA-N1/N7)]2+ complexes differ from the parent M(PMEA) complexes considerably; in the latter instance the formation of the five-membered chelates is of significance for all divalent metal ions studied. The observation that divalent metal-ion binding to the phosphonate group of (Dien)Pt(PMEA-N1) and (Dien)Pt(PMEA-N7) is only moderately inhibited (about 0.2-0.4 log units) by the twofold positively charged (Dien)Pt2+ unit at the adenine residue allows the general conclusion, considering that PMEA is a nucleotide analogue, that this is also true for nucleotides and that consequently participation of, for example, two metal ions in an enzymatic process involving nucleotides is not seriously hampered by charge repulsion.

Adenine↗

Synthesis, structure and spectral and redox properties of new mixed ligand monomeric and dimeric Ru(II) complexes: predominant formation of the "cis-alpha" diastereoisomer and unusual 3MC emission by dimeric complexes.

The tetradentate ligands 1,8-bis(pyrid-2-yl)-3,6-dithiaoctane (pdto) and 1,8-bis(benzimidazol-2-yl)-3,6-dithiaoctane (bbdo) form the complexes [Ru(pdto)(mu-Cl)](2)(ClO(4))(2) 1 and [Ru(bbdo)(mu-Cl)](2)(ClO(4))(2) 2 respectively. The new di-mu-chloro dimers 1 and 2 undergo facile symmetrical bridge cleavage reactions with the diimine ligands 2,2'-bipyridine (bpy) and dipyridylamine (dpa) to form the six-coordinate complexes [Ru(pdto)(bpy)](ClO(4))(2) 3, [Ru(bbdo)(bpy)](ClO(4))(2) 4, [Ru(pdto)(dpa)](ClO(4))(2) 5 and [Ru(bbdo)(dpa)](ClO(4))(2) 6 and with the triimine ligand 2,2':6,2''-terpyridine (terpy) to form the unusual seven-coordinate complexes [Ru(pdto)(terpy)](ClO(4))(2) 7 and [Ru(bbdo)(terpy)](ClO(4))(2) 8. In 1 the dimeric cation [Ru(pdto)(mu-Cl)](2)(2+) is made up of two approximately octahedrally coordinated Ru(II) centers bridged by two chloride ions, which constitute a common edge between the two Ru(II) octahedra. Each ruthenium is coordinated also to two pyridine nitrogen and two thioether sulfur atoms of the tetradentate ligand. The ligand pdto is folded around Ru(II) as a result of the cis-dichloro coordination, which corresponds to a "cis-alpha" configuration [DeltaDelta/LambdaLambda(rac) diastereoisomer] supporting the possibility of some attractive pi-stacking interactions between the parallel py rings at each ruthenium atom. The ruthenium atom in the complex cations 3a and 4 exhibit a distorted octahedral coordination geometry composed of two nitrogen atoms of the bpy and the two thioether sulfur and two py/bzim nitrogen atoms of the pdto/bbdo ligand, which is actually folded around Ru(II) to give a "cis-alpha" isomer. The molecule of complex 5 contains a six-coordinated ruthenium atom chelated by pdto and dpa ligands in the expected distorted octahedral fashion. The (1)H and (13)C NMR spectral data of the complexes throw light on the nature of metal-ligand bonding and the conformations of the chelate rings, which indicates that the dithioether ligands maintain their tendency to fold themselves even in solution. The bis-mu-chloro dimers 1 and 2 show a spin-allowed but Laporte-forbidden t(2g)(6)((1)A(1g))--> t(2g)(5) e(g)(1)((1)T(1g), (1)T(2g)) d-d transition. They also display an intense Ru(II) dpi--> py/bzim (pi*) metal-to-ligand charge transfer (MLCT) transition. The mononuclear complexes 3-8 exhibit dpi-->pi* MLCT transitions in the range 340-450 nm. The binuclear complexes 1 and 2 exhibit a ligand field ((3)MC) luminescence even at room temperature, whereas the mononuclear complexes 3 and 4 show a ligand based radical anion ((3)MLCT) luminescence. The binuclear complexes 1 and 2 undergo two successive oxidation processes corresponding to successive Ru(II)/Ru(III) couples, affording a stable mixed-valence Ru(II)Ru(III) state (K(c): 1, 3.97 x 10(6); 2, 1.10 x 10(6)). The mononuclear complexes 3-7 exhibit only one while 8 shows two quasi-reversible metal-based oxidative processes. The coordinated 'soft' thioether raises the redox potentials significantly by stabilising the 'soft' Ru(II) oxidation state. One or two ligand-based reduction processes were also observed for the mononuclear complexes.

Journal Article↗

Raman spectra of ionic liquids: a simulation study of AlF3 and its mixtures with NaF.

Theoretical Raman spectra of the melts of NaF/AlF3 mixtures have been obtained from computer simulations in order to examine how the Raman spectra reflect the coordination structure around the Al3+ ions. The Raman spectra, both polarized and depolarized, are calculated from a model for the dependence of the polarizability of the system on the ionic coordinates which was inspired by electronic structure calculations of the polarizabilities of ions in a condensed-phase environment. The shapes of the spectra and their evolution with composition in the mixtures conform remarkably well to those seen experimentally, and we discuss the relationship between the bands seen in the spectra and the vibrational modes of the AlFn(3-n) coordination complexes which are found in the melts. Finally, we calculate quantities which relate to the degree of cross-linking between these coordination complexes and their lifetimes.

Journal Article↗

Synthesis and magnetic properties of heterometal cyclic tetranuclear complexes [Cu(II)LM(II)(hfac)]2 (M(II) = Zn, Cu, Ni, Co, Fe, Mn; H3L = 1-(2-hydroxybenzamido)-2-((2-hydroxy-3-methoxybenzylidene)amino)ethane; Hhfac = hexafluoroacetylacetone).

A series of heterometal cyclic tetranuclear complexes [Cu(II)LM(II)(hfac)](2) (M(II) = Zn (1), Cu (2), Ni (3), Co (4), Fe(5), and Mn (6)) have been synthesized by the assembly reaction of K[CuL] and [M(II)(hfac)(2)(H(2)O)(2)] with a 1:1 mole ratio in methanol, where H(3)L = 1-(2-hydroxybenzamido)-2-((2-hydroxy-3-methoxybenzylidene)amino)ethane and Hhfac = hexafluoroacetylacetone. The crystal structures of 2, 4, and [Cu(II)LMn(II)(acac)](2) (6a) (Hacac = acetylacetone) were determined by single-crystal X-ray analyses. Each complex has a cyclic tetranuclear Cu(II)(2)M(II)(2) structure, in which the Cu(II) complex functions as a "bridging ligand complex", and the Cu(II) and M(II) ions are alternately arrayed. One side of the planar Cu(II) complex coordinates to one M(II) ion at the two phenoxo and the methoxy oxygen atoms, and the opposite side of the Cu(II) complex coordinates to another M(II) ion at the amido oxygen atom. The temperature-dependent magnetic susceptibilities revealed spin states of S(M) = 0, 1/2, 1, 3/2, 2, and 5/2 for the Zn(II), Cu(II), Ni(II), Co(II), Fe(II), and Mn(II) ions, respectively. Satisfactory fittings to the observed magnetic susceptibility data were obtained by assuming a rectangular arrangement with two different g-factors for the Cu(II) and M(II) ions, two different isotropic magnetic exchange interactions, J(1) and J(2), between the Cu(II) and M(II) ions, and a zero-field splitting term for the M(II) ion. In all cases, the antiferromagnetic coupling constants were found for both exchange interactions suggesting nonzero spin ground states with S(T) = 2/S(M) - S(Cu)/, which were confirmed by the analysis of the field-dependent magnetization measurements.

Journal Article↗

The ferrous heme of soluble guanylate cyclase: formation of hexacoordinate complexes with carbon monoxide and nitrosomethane.

The soluble form of guanylate cyclase (sGC) is the only definitive receptor for the signaling agent nitric oxide (.NO). The enzyme is a homologous heterodimer in which each subunit binds 1 equiv of 5-coordinate high-spin heme. .NO increases the Vmax of sGC up to 400-fold, probably by binding directly to the heme. Carbon monoxide (CO) forms a 6-coordinate complex with the heme and weakly activates the enzyme. Using stopped-flow spectrophotometry, the on-rate and off-rate for the binding of CO to the heme have been determined to be (3.58 +/- 0.15) x 10(4) M-1 s-1 and 3.5 +/- 0.5 s-1, respectively, at 10 degrees C. The equilibrium dissociation constant (Kd) has been independently determined to be 97 +/- 9 microM. Comparison of this Kd with that calculated from the rate constants indicates that the binding of CO to sGC is a simple one-step process, in which the off-rate of CO from the hexacoordinate complex is much faster than typically found in hemoproteins. The Kd of CO for activating the enzyme was also determined and compared to that for binding to the heme. Nitrosomethane forms irreversible complexes with typical ferrous hemoproteins but was observed to bind reversibly to the heme in sGC, with an off-rate > or = (7.6 +/- 0.2) x 10(-3) s-1. In general, the ferrous heme in sGC has a low affinity for ligands that form 6-coordinate complexes due primarily to fast ligand off-rates.

Animals↗