Biomedical subjects
Publications and source records attributed to M Porcelli.
A rapid, sensitive and specific high performance liquid chromatographic method for the simultaneous separation of natural adenosyl sulfur compounds has been developed. The method can be applied to biological samples for the estimation of S-adenosyl-L-methionine, S-adenosyl-L-homocysteine and S-adenosyl-(5')-3-methylthiopropylamine.
The paper reports the preparation and the chromatographic separation of new deaminated analogs of S-adenosyl(5')-3-methylthiopropylamine, i.e. S-adenosyl(5')-3-methylthiopropanol and S-inosyl(5')(-3-methylthiopropanol. These compounds can be used as specific inhibitors of polyamine biosynthesis. It is also reported the characterization of new sulfonium compounds by U.V. spectrophotometry, thin layer chromatography, high voltage electrophoresis, hydrolysis to known fragments.
5'-Methylthioadenosine phosphorylase has been purified approximately 340-fold in 20% yield from human prostate: the use of affinity chromatography by Sepharose-Hg has been found particularly advantageous. The enzyme has been partially characterized and an apparent Km of 2.5 x 10(-5) M has been calculated for 5'-methylthioadenosine. The reaction is activated by thiols and shows an absolute requirement for phosphate ions.
The uptake of labeled S-adenosylmethionine (SAM) by isolated and perfused rat liver has been compared to that of methionine: the rate of incorporation of the amino acid exceeds by about three times that of the sulfonium compound. S-Adenosylmethionine transport system shows saturation kinetics with an apparent Km of 59,5 microM and is energy-dependent, as demonstrated by the inhibition by 2,4-dinitrophenol.
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