Determination of argininosuccinate lyase and arginase activities with an amino acid analyzer

Antonio Bastone, Luisa Diomede, Rossella Parini, Franco Carnevale, Mario Salmona

Research output: Contribution to journalArticle

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Abstract

The measurement of argininosuccinate lyase (ASase) and arginase, both in liver and erythrocytes, was developed by using a commercial amino acid analyzer. The method is based upon the use of two different substrates, argininosuccinate and arginine for ASase and arginase, respectively, and the measurement of only one final metabolite: ornithine. The use of ornithine as a marker of biological activity of ASase is related to the fact that in the urea cycle, the specific activity of arginase is much higher than that of ASase; thus, during in vitro determinations, arginine, which is the product of ASase, is rapidly converted to ornithine. The sensitivity of the methods is very high since we were able to detect both activities using very diluted rat liver homogenates (0.10 mg protein/ml) or few microliters of human blood. In rat liver the Vmax for ASase and arginase were respectively 0.54 and 140 μmol/h/mg protein; the apparent Km values 1.25 and 13.5 mm. In human erythrocytes the Vmax for the same enzymes were 7.2 and 170 nmol/h/mg Hb and the apparent Km values were 0.66 and 9.5 mm. In 10 healthy volunteers the specific activity of ASase and arginase determined in blood were respectively 8.60 ± 0.46 and 124.1 ± 14.5 nmol/h/mg Hb. The results obtained from 2 patients suffering from argininosuccinic aciduria were also reported. In these latter cases while ASase was not detectable in blood, arginase activity was at the lowest end of the confidence limits determined in healthy volunteers.

Original languageEnglish
Pages (from-to)384-389
Number of pages6
JournalAnalytical Biochemistry
Volume191
Issue number2
DOIs
Publication statusPublished - 1990

Fingerprint

Argininosuccinate Lyase
Arginase
Amino Acids
Ornithine
Liver
Blood
Arginine
Rats
Argininosuccinic Aciduria
Healthy Volunteers
Erythrocytes
Activity Cycles
Metabolites
Bioactivity
Urea
Proteins
Biomarkers

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Determination of argininosuccinate lyase and arginase activities with an amino acid analyzer. / Bastone, Antonio; Diomede, Luisa; Parini, Rossella; Carnevale, Franco; Salmona, Mario.

In: Analytical Biochemistry, Vol. 191, No. 2, 1990, p. 384-389.

Research output: Contribution to journalArticle

Bastone, Antonio ; Diomede, Luisa ; Parini, Rossella ; Carnevale, Franco ; Salmona, Mario. / Determination of argininosuccinate lyase and arginase activities with an amino acid analyzer. In: Analytical Biochemistry. 1990 ; Vol. 191, No. 2. pp. 384-389.
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