Simona Porcu

Tyramine oxidation by copper/TPQ amine oxidase and peroxidase from Euphorbia characias latex

Tyramine, an important plant intermediate, was found to be a substrate for two proteins, a copper amine oxidase and a peroxidase from Euphorbia characias latex. The oxidation of tyramine took place by two different mechanisms: oxidative deamination to p-hydroxyphenylacetaldehyde by the amine oxidase and formation of di-tyramine by the peroxidase. The di-tyramine was further oxidized at the two amino groups by the amino oxidase, whereas p-hydroxyphenylacetaldehyde was transformed to di-p-hydroxyphenylacetaldehyde by the peroxidase. Data obtained in this study indicate a new interesting scenario in the metabolism of tyramine.

Some views on proteomics in diabetes

Abstract Mass spectrometry has been widely used in the field of diabetes. The development of new ionization methods and the effective coupling of mass spectrometry with liquid chromatography have enabled the protein modifications due to glycation processes to be investigated. Matrix assisted laser desorption/ionization mass spectrometry (MALDI/MS) has been used to evaluate the degree of glycation of specific plasma proteins. In contrast, the classic proteomic approach has been used to identify glycation sites and condensed sugar modifications. The same methods have been applied to studies on urinary protein profiles, enabling changes due to the development of long-term, diabetes-induced nephropathy to be identified. Published studies demonstrate that mass spectrometry is an important analytical tool for monitoring diabetes, capable of providing physicians with a new, more complete view of the physiopathological changes occurring as the disease develops.

Mass Spectrometry for Diabetic Nephropathy Monitoring: New Effective Tools for Physicians

The main aim of diabetic nephropathy monitoring is to identify molecular markers, that is, to find changes occurring at metabolome and proteome levels indicative of the diseases development. The mass spectrometry methods available today have been successfully applied to this field. This paper provides a short description of the basic aspects of the mass spectrometric methods used for diabetic nephropathy monitoring, reporting and discussing the results obtained using different approaches.

Increase in urinary purines and pyrimidines in patients with methylmalonic aciduria combined with homocystinuria.

BACKGROUND Methylmalonic aciduria combined with homocystinuria (MMA-HC) is the biochemical trait of a metabolic disorder resulting from impaired conversion of dietary cobalamin (cbl, or vitamin B12) to its two metabolically active forms. Effects on urinary purine and pyrimidine levels have not been described for this condition. METHODS Urine samples were collected from three patients with methylmalonic aciduria combined with homocystinuria and from 70 healthy subjects. Urinary purine and pyrimidine levels were quantitated by the use of LC/UV-Vis and LC/ESI/MS. RESULTS Higher urine levels of pyrimidines were detected with both methods in patients compared to controls. CONCLUSION Methylmalonic aciduria with homocystinuria is due to deficiency of the enzyme, cobalamin reductase. The enzyme defect leads to altered hepatic metabolism, which appears to modify circulating pyrimidine levels.

Structural and Functional Characterization of a New Double Variant Haemoglobin (HbG-Philadelphia/Duarte α268Asn→Lysβ262Ala→Pro)

We report the first case of cosegregation of two haemoglobins (Hbs): HbG-Philadelphia [α68(E17)Asn → Lys] and HbDuarte [β62(E6)Ala → Pro]. The proband is a young patient heterozygous also for β°-thalassaemia. We detected exclusively two haemoglobin variants: HbDuarte and HbG-Philadelphia/Duarte. Functional study of the new double variant HbG-Philadelphia/Duarte exhibited an increase in oxygen affinity, with a slight decrease of cooperativity and Bohr effect. This functional behaviour is attributed to β62Ala → Pro instead of α68Asn → Lys substitution. Indeed, HbG-Philadelphia isolated in our laboratory from blood cells donor carrier for this variant is not affected by any functional modification, whereas purified Hb Duarte showed functional properties very similar to the double variant. NMR and MD simulation studies confirmed that the presence of Pro instead of Ala at the β62 position produces displacement of the E helix and modifications of the tertiary structure. The substitution α68(E17)Asn → Lys does not cause significant structural and dynamical modifications of the protein. A possible structure-based rational of substitution effects is suggested.

Catabolic pathways for arginine and methylated arginines by plant and mammalian copper amine oxidases

The oxidation of L-arginine, monomethyl L-arginine and asymmetric dimethyl arginine catalyzed by copper/TPQ-amine oxidases from lentil (Lens esculenta) seedlings and pig kidney was investigated by optical spectroscopy and HPLC analyses. L-Arginine and its methylated derivatives were shown to be poor substrates for both enzymes and were oxidized by an unusual mechanism yielding glutamate-5-semialdehyde, ammonia, urea and their derivatives as reaction products. These findings suggest that amine oxidases might represent an alternative metabolic pathway of the arginine and its methylated derivatives, yielding new metabolites like urea, methylurea and dimethylurea.