A faclle enzymatic method for the location of radioactivity associated with the carbons of l-aspartic acid, l-asparagine, and β-cyano-l-alanine

David A. Cooney, Hiremagalur N. Jayaram

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

A radiometric enzymatic technique has been devised for locating radioactivity in the carbon skeleton of l-aspartic acid, l-asparagine, and β-cyano-l-alanine. l-Asparaginase at demonstrably discriminant concentrations is used to hydrolyze l-asparagine and/or β-cyano-l-alanine to l-aspartic acid, which in turn is transaminated to oxaloacetic acid by l-glutamate oxaloacetate transaminase. The β-carboxyl group of oxaloacetate is detached by zinc ions, and the radiolabeled CO2 is collected in alkali after diffusion. The residual pyruvic acid is α-decarboxylated by pyruvate decarboxylase to CO2, which is collected in a second alkaline trap, and the other product of decarboxylation, acetaldehyde, diffuses into a separate semicarbazide trap. Carbons 4, 1, and 3 plus 2 are located in this order. This method is shown to be facile, sensitive, reliable, and applicable to samples of biological origin.

Original languageEnglish (US)
Pages (from-to)508-520
Number of pages13
JournalAnalytical Biochemistry
Volume89
Issue number2
DOIs
StatePublished - 1978
Externally publishedYes

Fingerprint

Oxaloacetic Acid
Asparagine
Radioactivity
Aspartic Acid
Alanine
Carbon
Pyruvate Decarboxylase
Asparaginase
Decarboxylation
Acetaldehyde
Alkalies
Transaminases
Pyruvic Acid
Skeleton
Zinc
Glutamic Acid
Ions

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

A faclle enzymatic method for the location of radioactivity associated with the carbons of l-aspartic acid, l-asparagine, and β-cyano-l-alanine. / Cooney, David A.; Jayaram, Hiremagalur N.

In: Analytical Biochemistry, Vol. 89, No. 2, 1978, p. 508-520.

Research output: Contribution to journalArticle

@article{8e4f8407b22e4739a58c96db039f9502,
title = "A faclle enzymatic method for the location of radioactivity associated with the carbons of l-aspartic acid, l-asparagine, and β-cyano-l-alanine",
abstract = "A radiometric enzymatic technique has been devised for locating radioactivity in the carbon skeleton of l-aspartic acid, l-asparagine, and β-cyano-l-alanine. l-Asparaginase at demonstrably discriminant concentrations is used to hydrolyze l-asparagine and/or β-cyano-l-alanine to l-aspartic acid, which in turn is transaminated to oxaloacetic acid by l-glutamate oxaloacetate transaminase. The β-carboxyl group of oxaloacetate is detached by zinc ions, and the radiolabeled CO2 is collected in alkali after diffusion. The residual pyruvic acid is α-decarboxylated by pyruvate decarboxylase to CO2, which is collected in a second alkaline trap, and the other product of decarboxylation, acetaldehyde, diffuses into a separate semicarbazide trap. Carbons 4, 1, and 3 plus 2 are located in this order. This method is shown to be facile, sensitive, reliable, and applicable to samples of biological origin.",
author = "Cooney, {David A.} and Jayaram, {Hiremagalur N.}",
year = "1978",
doi = "10.1016/0003-2697(78)90380-9",
language = "English (US)",
volume = "89",
pages = "508--520",
journal = "Analytical Biochemistry",
issn = "0003-2697",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - A faclle enzymatic method for the location of radioactivity associated with the carbons of l-aspartic acid, l-asparagine, and β-cyano-l-alanine

AU - Cooney, David A.

AU - Jayaram, Hiremagalur N.

PY - 1978

Y1 - 1978

N2 - A radiometric enzymatic technique has been devised for locating radioactivity in the carbon skeleton of l-aspartic acid, l-asparagine, and β-cyano-l-alanine. l-Asparaginase at demonstrably discriminant concentrations is used to hydrolyze l-asparagine and/or β-cyano-l-alanine to l-aspartic acid, which in turn is transaminated to oxaloacetic acid by l-glutamate oxaloacetate transaminase. The β-carboxyl group of oxaloacetate is detached by zinc ions, and the radiolabeled CO2 is collected in alkali after diffusion. The residual pyruvic acid is α-decarboxylated by pyruvate decarboxylase to CO2, which is collected in a second alkaline trap, and the other product of decarboxylation, acetaldehyde, diffuses into a separate semicarbazide trap. Carbons 4, 1, and 3 plus 2 are located in this order. This method is shown to be facile, sensitive, reliable, and applicable to samples of biological origin.

AB - A radiometric enzymatic technique has been devised for locating radioactivity in the carbon skeleton of l-aspartic acid, l-asparagine, and β-cyano-l-alanine. l-Asparaginase at demonstrably discriminant concentrations is used to hydrolyze l-asparagine and/or β-cyano-l-alanine to l-aspartic acid, which in turn is transaminated to oxaloacetic acid by l-glutamate oxaloacetate transaminase. The β-carboxyl group of oxaloacetate is detached by zinc ions, and the radiolabeled CO2 is collected in alkali after diffusion. The residual pyruvic acid is α-decarboxylated by pyruvate decarboxylase to CO2, which is collected in a second alkaline trap, and the other product of decarboxylation, acetaldehyde, diffuses into a separate semicarbazide trap. Carbons 4, 1, and 3 plus 2 are located in this order. This method is shown to be facile, sensitive, reliable, and applicable to samples of biological origin.

UR - http://www.scopus.com/inward/record.url?scp=0018118155&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0018118155&partnerID=8YFLogxK

U2 - 10.1016/0003-2697(78)90380-9

DO - 10.1016/0003-2697(78)90380-9

M3 - Article

C2 - 365017

AN - SCOPUS:0018118155

VL - 89

SP - 508

EP - 520

JO - Analytical Biochemistry

JF - Analytical Biochemistry

SN - 0003-2697

IS - 2

ER -