Investigation of Structure and Rate of Synthesis of Ornithine Decarboxylase Protein in Mouse Kidney

Lo Persson, James E. Seely, Anthony Pegg

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

An immunoblotting technique was used to study the forms of ornithine decarboxylase present in androgen-in-duced mouse kidney. Two forms were detected which differed slightly in isoelectric point but not in subunit molecular weight (~55 000). Both forms were enzymatically active and could be labeled by reaction with radioactive a-(difluoromethyl)-ornithine, an enzyme-activated irreversible inhibitor. On storage of crude kidney homogenates or partially purified preparations of ornithine decarboxylase, the enzyme protein was degraded to a smaller size (Mr ~ 53 000) without substantial loss of enzyme activity. The synthesis and degradation of ornithine decarboxylase protein were studied by labeling the protein by intraperitoneal injection of [35S]methionine and immunoprecipitation using both monoclonal and polyclonal antibodies. The fraction of total protein synthesis represented by renal ornithine decarboxylase was increased at least 25-fold by testosterone treatment of female mice and was found to be about 1.1% in the fully induced androgen-treated female. Both forms of the enzyme were rapidly labeled in vivo, and the immunoprecipitable ornithine decarboxylase protein was almost completely lost after 4-h exposure to cycloheximide, confirming directly the very rapid turnover of this enzyme. Treatment with 1,3-diaminopropane which is known to cause a great reduction in ornithine decarboxylase activity did not greatly selectively inhibit the synthesis of the enzyme. However, 1,3-diaminopropane did produce an increase in the rate of degradation of ornithine decarboxylase and a general reduction in protein synthesis. These two factors, therefore, appear to be responsible for the loss of ornithine decarboxylase activity and protein in response to 1,3-diaminopropane.

Original languageEnglish (US)
Pages (from-to)3777-3783
Number of pages7
JournalBiochemistry
Volume23
Issue number16
DOIs
StatePublished - Jan 1 1984

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Ornithine Decarboxylase
Kidney
Proteins
Enzymes
Androgens
Degradation
Ornithine
Isoelectric Point
Enzyme activity
Cycloheximide
Intraperitoneal Injections
Immunoprecipitation
Immunoblotting
Methionine
Labeling
Testosterone
Thermodynamic properties
Molecular Weight
Molecular weight
Monoclonal Antibodies

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

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title = "Investigation of Structure and Rate of Synthesis of Ornithine Decarboxylase Protein in Mouse Kidney",
abstract = "An immunoblotting technique was used to study the forms of ornithine decarboxylase present in androgen-in-duced mouse kidney. Two forms were detected which differed slightly in isoelectric point but not in subunit molecular weight (~55 000). Both forms were enzymatically active and could be labeled by reaction with radioactive a-(difluoromethyl)-ornithine, an enzyme-activated irreversible inhibitor. On storage of crude kidney homogenates or partially purified preparations of ornithine decarboxylase, the enzyme protein was degraded to a smaller size (Mr ~ 53 000) without substantial loss of enzyme activity. The synthesis and degradation of ornithine decarboxylase protein were studied by labeling the protein by intraperitoneal injection of [35S]methionine and immunoprecipitation using both monoclonal and polyclonal antibodies. The fraction of total protein synthesis represented by renal ornithine decarboxylase was increased at least 25-fold by testosterone treatment of female mice and was found to be about 1.1{\%} in the fully induced androgen-treated female. Both forms of the enzyme were rapidly labeled in vivo, and the immunoprecipitable ornithine decarboxylase protein was almost completely lost after 4-h exposure to cycloheximide, confirming directly the very rapid turnover of this enzyme. Treatment with 1,3-diaminopropane which is known to cause a great reduction in ornithine decarboxylase activity did not greatly selectively inhibit the synthesis of the enzyme. However, 1,3-diaminopropane did produce an increase in the rate of degradation of ornithine decarboxylase and a general reduction in protein synthesis. These two factors, therefore, appear to be responsible for the loss of ornithine decarboxylase activity and protein in response to 1,3-diaminopropane.",
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Investigation of Structure and Rate of Synthesis of Ornithine Decarboxylase Protein in Mouse Kidney. / Persson, Lo; Seely, James E.; Pegg, Anthony.

In: Biochemistry, Vol. 23, No. 16, 01.01.1984, p. 3777-3783.

Research output: Contribution to journalArticle

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AU - Seely, James E.

AU - Pegg, Anthony

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AB - An immunoblotting technique was used to study the forms of ornithine decarboxylase present in androgen-in-duced mouse kidney. Two forms were detected which differed slightly in isoelectric point but not in subunit molecular weight (~55 000). Both forms were enzymatically active and could be labeled by reaction with radioactive a-(difluoromethyl)-ornithine, an enzyme-activated irreversible inhibitor. On storage of crude kidney homogenates or partially purified preparations of ornithine decarboxylase, the enzyme protein was degraded to a smaller size (Mr ~ 53 000) without substantial loss of enzyme activity. The synthesis and degradation of ornithine decarboxylase protein were studied by labeling the protein by intraperitoneal injection of [35S]methionine and immunoprecipitation using both monoclonal and polyclonal antibodies. The fraction of total protein synthesis represented by renal ornithine decarboxylase was increased at least 25-fold by testosterone treatment of female mice and was found to be about 1.1% in the fully induced androgen-treated female. Both forms of the enzyme were rapidly labeled in vivo, and the immunoprecipitable ornithine decarboxylase protein was almost completely lost after 4-h exposure to cycloheximide, confirming directly the very rapid turnover of this enzyme. Treatment with 1,3-diaminopropane which is known to cause a great reduction in ornithine decarboxylase activity did not greatly selectively inhibit the synthesis of the enzyme. However, 1,3-diaminopropane did produce an increase in the rate of degradation of ornithine decarboxylase and a general reduction in protein synthesis. These two factors, therefore, appear to be responsible for the loss of ornithine decarboxylase activity and protein in response to 1,3-diaminopropane.

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