An Antibody Probe to Determine the Native Species of Glycinamide Ribonucleotide Transformylase in Chicken Liver

M. Young, R. Douglas Sammons, W. Thomas Mueller, Stephen Benkovic

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Antibody probes of Western blots [Renart, J., Reiser, J., & Stark, G. (1979) Proc. Natl. Acad. Sci. U.S.A. 76, 3116] of chicken liver homogenates under various conditions revealed that glycinamide ribonucleotide transformylase can be rapidly proteolyzed in such homogenates. These findings, along with molecular weight measurements by ul-tracentrifugation, identify the true form of glycinamide ribonucleotide transformylase as a monomeric protein of 117 000 daltons. This protein has been purified 400-fold in 44% yield from chicken liver in one step on an affinity column of 10-formyl-5,8-dideazafolate-Sepharose. Native glycinamide ribonucleotide transformylase retains full activity after proteolytic cleavage to a form (Mr 55 000) similar to fragments seen in the Western blot of the homogenates. This phenomenon may be responsible for the previous identification of glycinamide ribonucleotide (GAR) transformylase as a dimer of 55 000-dalton subunits. Similar analyses using antibodies to 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) transformylase [Mueller, W. T., & Benkovic, S. J. (1981) Biochemistry 20, 337] and trifunctional enzyme [Smith, G. K., Mueller, W. T., Wasserman, G. F., Taylor, W. D., & Benkovic, S. J. (1980) Biochemistry 19, 4313] confirm that these two proteins were isolated in their native forms.

Original languageEnglish (US)
Pages (from-to)3979-3986
Number of pages8
JournalBiochemistry
Volume23
Issue number17
DOIs
StatePublished - Jan 1 1984

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Phosphoribosylglycinamide Formyltransferase
Liver
Chickens
Biochemistry
Antibodies
Hydroxymethyl and Formyl Transferases
Western Blotting
Ribonucleotides
Proteins
Weighing
Dimers
Sepharose
Molecular Weight
Molecular weight
Enzymes

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Young, M. ; Sammons, R. Douglas ; Mueller, W. Thomas ; Benkovic, Stephen. / An Antibody Probe to Determine the Native Species of Glycinamide Ribonucleotide Transformylase in Chicken Liver. In: Biochemistry. 1984 ; Vol. 23, No. 17. pp. 3979-3986.
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An Antibody Probe to Determine the Native Species of Glycinamide Ribonucleotide Transformylase in Chicken Liver. / Young, M.; Sammons, R. Douglas; Mueller, W. Thomas; Benkovic, Stephen.

In: Biochemistry, Vol. 23, No. 17, 01.01.1984, p. 3979-3986.

Research output: Contribution to journalArticle

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