Glutamyl Residues Research Articles

Показатели крови перепелов при скармливании муки из высушенных личинок Lucilia Caesar

The aim of the research was to study the effect of compound feed with the addition of flour from dried larvae of the Lucilia Caesar fly on the morphological and biochemical parameters of the blood of young quails. The experiment was conducted on 160 Phoenix quail at the age of 10 days, of which two groups were formed according to the principle of balanced groups. The results of the tests showed that all the studied blood parameters of the experimental bird were within the physiological norm. When feeding compound feed with a protein supplement from flour of the Lucilia Caesar larva, the content of albumins in the blood serum of quails of the second group decreased by 2.0 g/l (p≤0.05)., globulins - by 1.0 g/l (p≤0.05), urea - by 0.24 mmol/l (p≤0.01). Probably, a decrease in albumins and urea in the blood of quails of the second group indicates that they are involved in protein synthesis. Gamma-glutamyltransferase (GGT) is an enzyme involved in amino acid metabolism. Catalyzes the transfer of a gamma - glutamyl residue from a gamma-glutamyl peptide to an amino acid, another peptide, or, during hydrolysis, to water. The activity of the enzyme in the blood increases in liver diseases. In our experiment, a decrease in the GGT content by 2.66 u/l in the blood of a bird receiving flour from the larvae of the Lucilia Caesar fly probably indicates an activation of amino acid synthesis in the body, which is quite consistent with the data of the average daily increments of the experimental bird. Lactate dehydrogenase (LDH) in the blood is an enzyme localized inside cells. The composition necessarily includes zinc ions. The main function is to catalyze the oxidation of lactic acid to pyruvate. In diseases accompanied by tissue damage and cell destruction, LDH activity in the blood increases. In this regard, it is an important marker of tissue destruction. According to our data, LDH levels in the blood of the experimental bird of the second group significantly decreased by 2909.75 mmol/l (p≤0.05), which indicates an improvement in the functional activity of the liver.

Real-Time 1H NMR reveals position and sequence dependences of amino acid isomerization in amyloid beta fragments in situ

Amino acid isomers and their biologically relevant functions have been found in a variety of peptides and proteins in a living system. It is challenging, however, to specify how such isomerization proceeds at each amino acid independently in situ. In this work, the amino acid isomerization was quantified simultaneously at multiple residues in real time, by using solution-state 1H NMR in situ. The ability of NMR to distinguish isomerization product and reactant was demonstrated at different positions of aspartyl (Asp) residues in amyloid beta (Aβ) 1–40 fragments, as well as the glutamyl (Glu) residue. This enabled us to compare the isomerization of Asp1, Asp7, and Asp23, and Glu3 systematically in various Aβ fragments, in terms of the kinetics of isomerization in real time. It was found that Asp1 and Asp7 were prone to convert to iso-aspartyl (isoAsp) form, although Asp23 and Glu3 showed limited reactivity. The analysis demonstrated the role of histidine (His) in regulating Asp isomerization; His6 and His13/14 showed an inhibitory effect on the progress of isomerization at Asp1 and Asp7, respectively, by forming Asp-His complex that serves to slow down the conversion from Asp to isoAsp.

Polyglutamylation: biology and analysis

Polyglutamylation is a posttranslational modification (PTM) that adds several glutamates on glutamate residues in the form of conjugated peptide chains by a family of enzymes known as polyglutamylases. Polyglutamylation is well documented in microtubules. Polyglutamylated microtubules consist of different α- and β-tubulin subunits with varied number of added glutamate residues. Kinetic control and catalytic rates of tubulin modification by polyglutamylases influence the polyglutamylation pattern of functional microtubules. The recent studies uncovered catalytic mechanisms of the glutamylation enzymes family, particularly tubulin tyrosine ligase-like (TTLL). Variable length polyglutamylation of primary sequence glutamyl residues have been mapped with a multitude of protein chemistry and proteomics approaches. Although polyglutamylation was initially considered a tubulin-specific modification, the recent studies have uncovered a calmodulin-dependent glutamylase, SidJ. Nano-electrospray ionization (ESI) proteomic approaches have identified quantifiable polyglutamylated sites in specific substrates. Indeed, conjugated glutamylated peptides were used in nano-liquid chromatography gradient delivery due to their relative hydrophobicity for their tandem mass spectrometry (MS/MS) characterization. The recent polyglutamylation characterization has revealed three major sites: E445 in α-tubulin, E435 in β-tubulin, and E860 in SdeA. In this review, we have summarized the progress made using proteomic approaches for large-scale detection of polyglutamylated peptides, including biology and analysis.

Molecular characterization of a novel aspartyl aminopeptidase that contributes to the increase in glutamic acid content in chicken meat during cooking

The enzyme involved in the increase in glutamic acid content in chicken meat during cooking was identified and characterized. Chicken homogenate produced significantly more free glutamic acid and exhibited higher glutamyl p-nitroanilide (Glu-pNA) hydrolyzing activity than beef when heat cooked. Amino acid sequencing revealed the presence of aspartyl aminopeptidase (DNPEP) in chicken meat. Using RT-PCR, DNPEP gene expression was detected in chicken breast and thigh muscles, liver, and small intestine, together with various other peptidase genes. Full-length DNPEP cDNA was cloned, and recombinant chicken DNPEP (cDNPEP) was expressed in Escherichia coli. cDNPEP showed five-fold higher activity against Glu-pNA than against aspartyl-pNA, which represents a different substrate specificity than observed for recombinant bovine DNPEP (bDNPEP). The Km values of both DNPEPs with Glu p-NA substrates indicated a higher affinity of cDNPEP for glutamyl residues. This unique substrate specificity of cDNPEP contributes to efficient glutamic acid production in chickens.

Structural basis for non-radical catalysis by TsrM, a radical SAM methylase.

TsrM methylates C2 of the indole ring of L-tryptophan (Trp) during the biosynthesis of the quinaldic acid moiety of thiostrepton. It is annotated as a cobalamin-dependent radical S-adenosylmethionine (SAM) methylase; however, TsrM does not reductively cleave SAM to the universal 5ʹ-deoxyadenosyl 5ʹ-radical intermediate, a hallmark of radical-SAM (RS) enzymes. Herein, we report structures of TsrM from Kitasatospora setae, the first of a cobalamin-dependent radical SAM methylase. Unexpectedly, the structures show an essential arginine residue that resides in the proximal coordination sphere of the cobalamin cofactor and a [4Fe–4S] cluster that is ligated by a glutamyl residue and three cysteines in a canonical CxxxCxxC RS motif. Structures in the presence of substrates suggest a substrate-assisted mechanism of catalysis, wherein the carboxylate group of SAM serves as a general base to deprotonate N1 of the tryptophan substrate, facilitating formation of a C2 carbanion.

Methyltransferase CheR binds to its chemoreceptor substrates independent of their signaling conformation yet modifies them differentially.

Methylation of specific chemoreceptor glutamyl residues by methyltransferase CheR mediates sensory adaptation and gradient sensing in bacterial chemotaxis. Enzyme action is a function of chemoreceptor signaling conformation: kinase-off receptors are more readily methylated than kinase-on, a feature central to adaptational and gradient-sensing mechanisms. Differential enzyme action could reflect differential binding, catalysis or both. We investigated by measuring CheR binding to kinase-off and kinase-on forms of Escherichia coli aspartate receptor Tar deleted of its CheR-tethering, carboxyl terminus pentapeptide. This allowed characterization of the low-affinity binding of enzyme to the substrate receptor body, otherwise masked by high-affinity interaction with pentapeptide. We quantified the low-affinity protein-protein interactions by determining kinetic rate constants of association and dissociation using bio-layer interferometry and from those values calculating equilibrium constants. Whether Tar signaling conformations were shifted by ligand occupancy or adaptational modification, there was little or no difference between the two signaling conformations in kinetic or equilibrium parameters of enzyme-receptor binding. Thus, differential methyltransferase action does not reflect differential binding. Instead, the predominant determinants of binding must be common to different signaling conformations. Characterization of the dependence of association rate constants on Deybe length, a measure of the influence of electrostatics, implicated electrostatic interactions as a common binding determinant. Taken together, our observations indicate that differential action of methyltransferase on kinase-off and kinase-on chemoreceptors is not the result of differential binding and suggest it reflects differential catalytic propensity. Differential catalysis rather than binding could well be central to other enzymes distinguishing alternative conformations of protein substrates.

Surface-exposed domains of TatB involved in the structural and functional assembly of the Tat translocase in Escherichia coli

Twin-arginine-dependent translocases transport folded proteins across bacterial, archaeal, and chloroplast membranes. Upon substrate binding, they assemble from hexahelical TatC and single-spanning TatA and TatB membrane proteins. Although structural and functional details of individual Tat subunits have been reported previously, the sequence and dynamics of Tat translocase assembly remain to be determined. Employing the zero-space cross-linker N,N'-dicyclohexylcarbodiimide (DCCD) in combination with LC-MS/MS, we identified as yet unknown intra- and intermolecular contact sites of TatB and TatC. In addition to their established intramembrane binding sites, both proteins were thus found to contact each other through the soluble N terminus of TatC and the interhelical linker region around the conserved glutamyl residue Glu49 of TatB from Escherichia coli Functional analyses suggested that by interacting with the TatC N terminus, TatB improves the formation of a proficient substrate recognition site of TatC. The Glu49 region of TatB was found also to contact distinct downstream sites of a neighboring TatB molecule and to thereby mediate oligomerization of TatB within the TatBC receptor complex. Finally, we show that global DCCD-mediated cross-linking of TatB and TatC in membrane vesicles or, alternatively, creating covalently linked TatC oligomers prevents TatA from occupying a position close to the TatBC-bound substrate. Collectively, our results are consistent with a circular arrangement of the TatB and TatC units within the TatBC receptor complex and with TatA entering the interior TatBC-binding cavity through lateral gates between TatBC protomers.

A New Horizon in Vitamin K Research.

Vitamin K is a cofactor for γ-glutamyl carboxylase, which catalyzes the posttranslational conversion of specific glutamyl residues to γ-carboxyglutamyl residues in a variety of vitamin K-dependent proteins (VKDPs) involved in blood coagulation, bone and cartilage metabolism, signal transduction, and cell proliferation. Despite the great advances in the genetic, structural, and functional studies of VKDPs as well as the enzymes identified as part of the vitamin K cycle which enable it to be repeatedly recycled within the cells, little is known of the identity and roles of key regulators of vitamin K metabolism in mammals and humans. This review focuses on new insights into the molecular mechanisms underlying the intestinal absorption and in vivo tissue conversion of vitamin K1 to menaquinone-4 (MK-4) with special emphasis on two major advances in the studies of intestinal vitamin K transporters in enterocytes and a tissue MK-4 biosynthetic enzyme UbiA prenyltransferase domain-containing protein 1 (UBIAD1), which participates in the in vivo conversion of a fraction of dietary vitamin K1 to MK-4 in mammals and humans, although it remains uncertain whether UBIAD1 functions as a key regulator of intracellular cholesterol metabolism, bladder and prostate tumor cell progression, vascular integrity, and protection from oxidative stress.

Preparation of phylloquinone-loaded poly(lactic acid)/hydroxyapatite core–shell particles and their drug release behavior

Poly(lactic acid)/hydroxyapatite (PLA/HAp) core–shell particles loaded with hydrophobic drugs were fabricated through a modified surfactant-free emulsification method. The core–shell particles are expected to encapsulate hydrophobic drugs, whose clinical use is often hampered because of their poor water solubility. In this study, phylloquinone, a drug that mediates the gamma carboxylation of glutamyl residues, was used as a model hydrophobic drug. Oil-in-water emulsions were prepared using an organic phase composed of dissolved PLA with various amounts of phylloquinone and an aqueous phase composed of calcium acetate and diammonium hydrogen phosphate. The obtained suspension exhibited a yellow color, indicating that the particles successfully incorporated phylloquinone. Scanning electron microscopy and X-ray diffraction analyses revealed these particles to be PLA/HAp core–shell composites loaded with phylloquinone, approximately 100–500nm in diameter. The particle diameters were affected by the amount of loaded phylloquinone; excess phylloquinone causes encapsulation failure. The drug release behavior of the phylloquinone-loaded PLA/HAp particles was examined via UV detection. The phylloquinone release profile showed a sustained release from the PLA/HAp core–shell particles over a ten days period.

Linked production of pyroglutamate-modified proteins via self-cleavage of fusion tags with TEV protease and autonomous N-terminal cyclization with glutaminyl cyclase in vivo.

Overproduction of N-terminal pyroglutamate (pGlu)-modified proteins utilizing Escherichia coli or eukaryotic cells is a challenging work owing to the fact that the recombinant proteins need to be recovered by proteolytic removal of fusion tags to expose the N-terminal glutaminyl or glutamyl residue, which is then converted into pGlu catalyzed by the enzyme glutaminyl cyclase. Herein we describe a new method for production of N-terminal pGlu-containing proteins in vivo via intracellular self-cleavage of fusion tags by tobacco etch virus (TEV) protease and then immediate N-terminal cyclization of passenger target proteins by a bacterial glutaminyl cyclase. To combine with the sticky-end PCR cloning strategy, this design allows the gene of target proteins to be efficiently inserted into the expression vector using two unique cloning sites (i.e., SnaB I and Xho I), and the soluble and N-terminal pGlu-containing proteins are then produced in vivo. Our method has been successfully applied to the production of pGlu-modified enhanced green fluorescence protein and monocyte chemoattractant proteins. This design will facilitate the production of protein drugs and drug target proteins that possess an N-terminal pGlu residue required for their physiological activities.

Purification and biochemical characterization of feruloyl esterases fromAspergillus terreusMTCC 11096

Aspergillus terreus MTCC 11096 isolated from the soils of agricultural fields cultivating sweet sorghum was previously identified to produce feruloyl esterases (FAEs). The enzymes responsible for feruloyl esterase activity were purified to homogeneity and named as AtFAE-1, AtFAE-2, and AtFAE-3. The enzymes were monomeric having molecular masses of 74, 23 and 36 kDa, respectively. Active protein bands were identified by a developed pH-dependent zymogram on native PAGE. The three enzymes exhibited variation in pH tolerance ranging between pH 5-8 and thermostability of up to 55°C. Inhibition studies revealed that the serine residue was essential for feruloyl esterase activity; moreover aspartyl and glutamyl residues are not totally involved at the active site. Metal ions such as Ca(2+), K(+), and Mg(2+) stabilized the enzyme activity for all three FAEs. Kinetic data indicated that all three enzymes showed catalytic efficiencies (k(cat) /K(m)) against different synthesized alkyl and aryl esters indicating their broad substrate specificity. The peptide mass fingerprinting by MALDI/TOF-MS analysis and enzyme affinity toward methoxy and hydroxy substituents on the benzene ring revealed that the AtFAE-1 belonged to type A while AtFAE-2 and AtFAE-3 were type C FAE. The FAEs could release 65 to 90% of ferulic acid from agrowaste substrates in the presence of xylanase.

Two glutamic acid residues in the DNA-binding domain are engaged in the release of STAT1 dimers from DNA

BackgroundIn interferon-γ-stimulated cells, the dimeric transcription factor STAT1 (signal transducer and activator of transcription 1) recognizes semi-palindromic motifs in the promoter regions of cytokine-driven target genes termed GAS (gamma-activated sites). However, the molecular steps that facilitate GAS binding and the subsequent liberation of STAT1 homodimers from these promoter elements are not well understood.ResultsUsing a mutational approach, we identified two critical glutamyl residues within the DNA-binding domain adjacent to the phosphodiester backbone of DNA which efficiently release phospho-STAT1 from DNA. The release of STAT1 dimers from DNA enhances transcriptional activity on both interferon-driven reporter and endogenous target genes. A substitution of either of the two glutamic acid residues broadens the repertoire of putative binding sites on DNA and enhances binding affinity to GAS sites. However, despite elevated levels of tyrosine phosphorylation and a prolonged nuclear accumulation period, the STAT1 DNA-binding mutants show a significantly reduced transcriptional activity upon stimulation of cells with interferon-γ. This reduced transcriptional response may be explained by the deposition of oligomerized STAT1 molecules outside GAS sites.ConclusionsThus, two negatively charged amino acid residues in the DNA-binding domain are engaged in the liberation of STAT1 from DNA, resulting in a high dissociation rate from non-GAS sites as a key feature of STAT1 signal transduction, which positively regulates cytokine-dependent gene expression probably by preventing retention at transcriptionally inert sites.

Intracellular Disposition of Methotrexate in Acute Lymphoblastic Leukemia in Children

Methotrexate (MTX) is a key agent for the treatment of acute lymphoblastic leukemia in children and the benefit of high-dose MTX is well established as it significantly increases cure rates and improves patients' prognosis. However, the determinants of MTX therapeutic effect are not clearly identified, although intracellular polyglutamation is essential. MTX, the monoglutamate form (MTXG₁) inhibits the dihydrofolate reductase (DHFR) implicated in the folate cycle. MTXG₁ is metabolized to active methotrexate polyglutamates (MTXPG) with sequential gamma-linkage of 2 to 6 glutamyl residues by the folylpolyglutamate synthetase (FPGS). Long chain MTXPG have higher affinity than MTX for the enzymes involved in de novo purine synthesis such as 5-aminoimidazole-4-carboxamide ribonucleotide transformylase (ATIC) and thymidilate synthase (TS), which results in a reinforcement of MTX inhibition. Thus, intracellular formation of MTXPG enhances the cytotoxic and antileukemic effect of MTX. Different pharmacogenetic polymorphisms contribute to interindividual variability in MTX response to treatment. In addition, pharmacokinetic interactions with 6-mercaptopurine (6-MP), frequently co-administered, have been reported. And factors affecting intracellular MTX disposition and 6-MP/MTX interactions, including pharmacogenetic polymorphisms affecting MTX disposition are reviewed.

Aggregation of glutathione-functionalized Au nanoparticles induced by Ni2+ ions

Aggregation of glutathione (GSH)-functionalized Au nanoparticles induced by Ni2+ ions were found to be related to pH of the solutions. At pH lower than 9.0, introduction of Ni2+ ions was less effective to induce aggregation of the Au nanoparticles. At pH around 9.8, the Au nanoparticles experienced extensive aggregation upon the addition of Ni2+ ions. When pH was higher than 10.5, Ni2+ ion-induced aggregation of the Au nanoparticles was suppressed gradually with increasing pH. It was identified that such pH-mediated aggregation behaviors are attributed to the different coordination fashions of GSH on the Au nanoparticle surface with Ni2+ ions. At pH lower than 9.0, addition of Ni2+ ions was less effective to induce aggregation of the Au nanoparticles when only the carboxyl group of the glutamyl residue was available for the metal ions. The Au nanoparticles underwent extensive aggregation at pH around 9.8 when both the carboxyl and deprotonated amine groups of the glutamyl residue in GSH could coordinate with the metal ions. At pH higher than 10.5, the coordination was gradually suppressed by the hydroxyl groups in solutions, addition of Ni2+ ions is less effective to cross-link the Au nanoparticles. Such a work is helpful for understanding the sensitivity and selectivity of GSH-functionalized Au nanoparticles to metal ions.

Glutamate: a truly functional amino acid

Glutamate is one of the most abundant of the amino acids. In addition to its role in protein structure, it plays critical roles in nutrition, metabolism and signaling. Post-translational carboxylation of glutamyl residues increases their affinity for calcium and plays a major role in hemostasis. Glutamate is of fundamental importance to amino acid metabolism, yet the great bulk of dietary glutamate is catabolyzed within the intestine. It is necessary for the synthesis of key molecules, such as glutathione and the polyglutamated folate cofactors. It plays a major role in signaling. Within the central nervous system, glutamate is the major excitatory neurotransmitter and its product, GABA, the major inhibitory neurotransmitter. Glutamate interaction with specific taste cells in the tongue is a major component of umami taste. The finding of glutamate receptors throughout the gastrointestinal tract has opened up a new vista in glutamate function. Glutamate is truly a functional amino acid.

γ-Glutamylamines and neurodegenerative diseases

Transglutaminases catalyze the formation of γ-glutamylamines utilizing glutamyl residues and amine-bearing compounds such as lysyl residues and polyamines. These γ-glutamylamines can be released from proteins by proteases in an intact form. The free γ-glutamylamines can be catabolized to 5-oxo-L-proline and the free amine by γ-glutamylamine cyclotransferase. Free γ-glutamylamines, however, accumulate in the CSF and affected areas of Huntington Disease brain. This observation suggests transglutaminase-derived γ-glutamylamines may play a more significant role in neurodegeneration than previously thought. The following monograph reviews the metabolism of γ-glutamylamines and examines the possibility that these species contribute to neurodegeneration.

Female distance runners show a different response to post-workout consumption of wheat gluten hydrolysate compared to their male counterparts

Wheat gluten hydrolysate (WGH) is rich in glutamyl residue; glutamine is considered a conditionally essential amino acid under physical stress. WGH has been reported to suppress post-exercise rises in serum creatine kinase in male distance runners. This study aimed to reproduce the effects in female distance runners under similar conditions. The study was conducted in a double-blinded crossover manner. Six female collegiate distance runners ingested WGH or a placebo after a 2-h run at an intensity estimated as 60-70% of their maximum oxygen uptake. Blood was sampled before, immediately after, and at 10 and 24 h after the run. Unlike those in male runners, serum creatine kinase (CK) increased slightly, with a peak at 10 h after the run, while plasma glutamine kept declining. The anti-inflammatory effect of WGH was not evident since the post-exercise elevation of CK was ambiguous. Plasma glutamine concentrations also showed a different kinetics from that in men.

Direct evidence that the carboxyl‐terminal sequence of a bacterial chemoreceptor is an unstructured linker and enzyme tether

Sensory adaptation in bacterial chemotaxis involves reversible methylation of specific glutamyl residues on chemoreceptors. The reactions are catalyzed by a dedicated methyltransferase and dedicated methylesterase. In Escherichia coli and related organisms, control of these enzymes includes an evolutionarily recent addition of interaction with a pentapeptide activator located at the carboxyl terminus of the receptor polypeptide chain. Effective enzyme activation requires not only the pentapeptide but also a segment of the receptor polypeptide chain between that sequence and the coiled-coil body of the chemoreceptor. This segment has features consistent with a role as a flexible and presumably unstructured linker and enzyme tether, but there has been no direct information about its structure. We used site-directed spin labeling and electron paramagnetic resonance spectroscopy to characterize structural features of the carboxyl-terminal 40 residues of E. coli chemoreceptor Tar. Beginning ∼ 35 residues from the carboxyl terminus and continuing to the end of the protein, spectra of spin-labeled Tar embedded in native membranes or in reconstituted proteoliposomes, exhibited mobilities characteristic of unstructured, disordered segments. Binding of methyltransferase substantially reduced mobility for positions in or near the pentapeptide but mobility for the linker sequence remained high, being only modestly reduced in a gradient of decreasing effects for 10-15 residues, a pattern consistent with the linker providing a flexible arm that would allow enzyme diffusion within defined limits. Thus, our data identify that the carboxyl-terminal linker between the receptor body and the pentapeptide is an unstructured, disordered segment that can serve as a flexible arm and enzyme tether.

Optical resolution of racemic amino acid derivatives with chiral polyamides bearing glutamyl residue as a diacid component

AbstractNovel chiral polyamides with chiral environment in their main chains were obtained from aromatic diamine, 4,4′‐diaminodiphenylmethane (DADPM), and the D‐isomer or the L‐isomer of N‐α‐protected glutamic acid, such as N‐α‐benzyloxycarbonyl‐D‐glutamic acid (Z‐D‐Glu‐OH) or N‐α‐benzyloxycarbonyl‐L‐glutamic acid (Z‐L‐Glu‐OH), in the presence of triphenyl phosphite (TPP). Two types of newly prepared polyamide showed optical rotation, implying that there were asymmetric carbons in their main chains. Circular dichroism studies demonstrated that resulting chiral poly‐ amides took a helical structure. Optical resolution ability of those two types of polyamide was studied by adopting potential difference as a driving force for membrane transport. Membranes showed permselectivity toward racemic mixture of N‐α‐acetyltryptophan (Ac‐Trp). The permselectivity was dependent on the absolute configuration of diacid component. The permselectivity was expressed by diffusivity selectivity, which was determined by the presence of chiral helicity. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2011

Palladium(II) complexes of oligopeptides containing aspartyl and glutamyl residues

Potentiometric and 1H NMR spectroscopic measurements have been performed on palladium(II) complexes of di-, tri- and tetra-peptides containing aspartyl and glutamyl residues including AspAla, AlaAsp, AspAsp, AspAspAsp, GluGluGlu, GlyAspGly, AspAspAspAsp and GlyGlyAspGly. In the case of dipeptides the coordination modes are basically determined by the peptide backbone. The presence of the extra β-carboxylate residues does not result in new binding modes but these functions may slightly affect the thermodynamic stability of several species. For tripeptides the most important findings are connected to the governing role of the β-carboxylate group of internal aspartyl residues but similar effects were not observed for the peptides containing glutamic acid. Aspartic acid in the second position of a tripeptide (Xaa-Asp-Yaa sequence) promotes the binding of the preceding and prevents deprotonation and coordination of subsequent amide functions. In the case of tetrapeptides, the aspartyl residues present in the third position from the N-terminus (Xaa-Yaa-Asp-Zaa sequence) have the most pronounced effect on complex formation. In this case, the (NH 2, N −, N −, β-COO −)-coordination is the major binding mode and the species [PdH −2L] can exist in a wide pH range. The enhanced stability of these complexes was explained by the conformational changes of the coordinated ligands.