Gamma glutamyl transpeptidase: Measurement and development in guinea pig small intestine

Michael I. Cohen, Lawrence M. Gartner, Olga O. Blumenfeld, Irwin M. Arias

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Extract: A modification of a previous method for the assay of γ glutamyl transpeptidase (GGTP) was developed. Substrate solubility difficulties alluded to by other investigators were avoided by employing heating and solubilization of the chromogenic substrate γ glutamyl-β-naphthylamide in a medium of carbonate 0.05 M and Tris buffer 0.1 M at pH 9.5. The kinetics and conditions for such an assay are described. Whole intestinal homogenates of adult male guinea pigs were used as the source of the enzyme. The developmental pattern of this enzyme was determined in fetuses at 55 and 63 days of gestation and at varying times from 1 to 90 days of age. A total of 69 animals was assayed. The general pattern was that of high specific activity during prenatal life, with a rapid decline during the neonatal period (3–24 days of age) and a slight increase after 55 days of age. Other guinea pig organs were studied. Liver and kidney were found to contain enzyme activity greater than that of the intestine. Subcellular fractionation of intestinal mucosa using ultracentrifugation revealed a twenty-fold enrichment of activity in jejunal brush border membrane, compared with whole jejunal homogenates when expressed as specific enzyme activity per mg of protein. The stability characteristics of GGTP disclosed no loss of specific activity when stored at−28° for 50 days. This simple enzyme assay, stability of the enzyme when frozen, subcellular distribution in the intestinal brush border membrane, and an unusual developmental pattern made this enzyme a useful adjunct to the study of intestinal protein metabolism. Speculation: The unique ability of GGTP to hydrolyze > glutamic acid-peptide bonds and the location of the enzyme in the intestinal brush border suggest a role for this enzyme in the metabolism of an unusual group of physiologically important peptides such as glutathione, folic acid, and gluten-gliadin. The significance of these γ-bonded compounds may now be approached in order to investigate the role of this enzyme in gluten enteropathy and related disorders.

Original languageEnglish (US)
Pages (from-to)5-10
Number of pages6
JournalPediatric Research
Volume3
Issue number1
StatePublished - 1969

Fingerprint

gamma-Glutamyltransferase
Small Intestine
Guinea Pigs
Enzymes
Microvilli
Chromogenic Compounds
Gliadin
Enzyme Stability
Peptides
Tromethamine
Membranes
Glutens
Ultracentrifugation
Carbonates
Enzyme Assays
Celiac Disease
Intestinal Mucosa
Folic Acid
Solubility
Heating

Keywords

  • Biochemistry
  • Developmental
  • Glutamyl transpeptidase
  • Intestine
  • Kidney
  • Liver

ASJC Scopus subject areas

  • Pediatrics, Perinatology, and Child Health

Cite this

Gamma glutamyl transpeptidase : Measurement and development in guinea pig small intestine. / Cohen, Michael I.; Gartner, Lawrence M.; Blumenfeld, Olga O.; Arias, Irwin M.

In: Pediatric Research, Vol. 3, No. 1, 1969, p. 5-10.

Research output: Contribution to journalArticle

Cohen, MI, Gartner, LM, Blumenfeld, OO & Arias, IM 1969, 'Gamma glutamyl transpeptidase: Measurement and development in guinea pig small intestine', Pediatric Research, vol. 3, no. 1, pp. 5-10.
Cohen, Michael I. ; Gartner, Lawrence M. ; Blumenfeld, Olga O. ; Arias, Irwin M. / Gamma glutamyl transpeptidase : Measurement and development in guinea pig small intestine. In: Pediatric Research. 1969 ; Vol. 3, No. 1. pp. 5-10.
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N2 - Extract: A modification of a previous method for the assay of γ glutamyl transpeptidase (GGTP) was developed. Substrate solubility difficulties alluded to by other investigators were avoided by employing heating and solubilization of the chromogenic substrate γ glutamyl-β-naphthylamide in a medium of carbonate 0.05 M and Tris buffer 0.1 M at pH 9.5. The kinetics and conditions for such an assay are described. Whole intestinal homogenates of adult male guinea pigs were used as the source of the enzyme. The developmental pattern of this enzyme was determined in fetuses at 55 and 63 days of gestation and at varying times from 1 to 90 days of age. A total of 69 animals was assayed. The general pattern was that of high specific activity during prenatal life, with a rapid decline during the neonatal period (3–24 days of age) and a slight increase after 55 days of age. Other guinea pig organs were studied. Liver and kidney were found to contain enzyme activity greater than that of the intestine. Subcellular fractionation of intestinal mucosa using ultracentrifugation revealed a twenty-fold enrichment of activity in jejunal brush border membrane, compared with whole jejunal homogenates when expressed as specific enzyme activity per mg of protein. The stability characteristics of GGTP disclosed no loss of specific activity when stored at−28° for 50 days. This simple enzyme assay, stability of the enzyme when frozen, subcellular distribution in the intestinal brush border membrane, and an unusual developmental pattern made this enzyme a useful adjunct to the study of intestinal protein metabolism. Speculation: The unique ability of GGTP to hydrolyze > glutamic acid-peptide bonds and the location of the enzyme in the intestinal brush border suggest a role for this enzyme in the metabolism of an unusual group of physiologically important peptides such as glutathione, folic acid, and gluten-gliadin. The significance of these γ-bonded compounds may now be approached in order to investigate the role of this enzyme in gluten enteropathy and related disorders.

AB - Extract: A modification of a previous method for the assay of γ glutamyl transpeptidase (GGTP) was developed. Substrate solubility difficulties alluded to by other investigators were avoided by employing heating and solubilization of the chromogenic substrate γ glutamyl-β-naphthylamide in a medium of carbonate 0.05 M and Tris buffer 0.1 M at pH 9.5. The kinetics and conditions for such an assay are described. Whole intestinal homogenates of adult male guinea pigs were used as the source of the enzyme. The developmental pattern of this enzyme was determined in fetuses at 55 and 63 days of gestation and at varying times from 1 to 90 days of age. A total of 69 animals was assayed. The general pattern was that of high specific activity during prenatal life, with a rapid decline during the neonatal period (3–24 days of age) and a slight increase after 55 days of age. Other guinea pig organs were studied. Liver and kidney were found to contain enzyme activity greater than that of the intestine. Subcellular fractionation of intestinal mucosa using ultracentrifugation revealed a twenty-fold enrichment of activity in jejunal brush border membrane, compared with whole jejunal homogenates when expressed as specific enzyme activity per mg of protein. The stability characteristics of GGTP disclosed no loss of specific activity when stored at−28° for 50 days. This simple enzyme assay, stability of the enzyme when frozen, subcellular distribution in the intestinal brush border membrane, and an unusual developmental pattern made this enzyme a useful adjunct to the study of intestinal protein metabolism. Speculation: The unique ability of GGTP to hydrolyze > glutamic acid-peptide bonds and the location of the enzyme in the intestinal brush border suggest a role for this enzyme in the metabolism of an unusual group of physiologically important peptides such as glutathione, folic acid, and gluten-gliadin. The significance of these γ-bonded compounds may now be approached in order to investigate the role of this enzyme in gluten enteropathy and related disorders.

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