The covalent attachment of polyamines to proteins in plant mitochondria

T V Votyakova, H M Wallace, B Dunbar, S B Wilson

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Plant mitochondria from both potato end mung bean incorporated radioactivity into acid insoluble material when incubated with labelled polyamines (spermine, spermidine and putrescine). Extensive washing of mitochondrial precipitates with trichloroncetic acid and the excess of cold polyamine failed to remove bound radioactivity. Addition of nonradioactive polyamine stopped further incorporation of radioactivity but did net release radioactivity already bound. The radioactivity is incorporated into the membrane fraction, The labelling process has all the features of an enzymatic reaction: it is long lasting with distinctive kinetics peculiar to each polyamine, it is temperature dependent and is affected by N-ethylmaleimide, The latter inhibits the incorporation of putrescine but stimulates the incorporation of spermine and spermidine. Treatment of prelabelled mitochondria with pepsin releases bound radioactivity thus indicating protein to be the ligand for the attachment of polyamines. HPLC of mitochondrial hydrolysates revealed that the radioactivity bound to mitochondria is polyamines; traces of acetyl polyamines were also found in some samples. On autoradiograms of SDS/PAGE gels several radioactive bands of proteins were detected. Protein sequencing of labelled spots from a 2D gel gave a sequence which was 60% identical to catalase. We suggest that the attachment of polyamines to mitochondrial proteins occurs cotranslationally possibly via transglutaminases.

Original languageEnglish
Pages (from-to)250-257
Number of pages8
JournalEuropean Journal of Biochemistry
Volume260
Publication statusPublished - 1999

Keywords

  • plant mitochondria
  • polyamines
  • covalent modification
  • protein modification
  • catalase
  • HELIANTHUS-TUBEROSUS
  • SPERMINE TRANSPORT
  • TRANSGLUTAMINASES
  • IDENTIFICATION
  • DERIVATIVES
  • MEMBRANES
  • APOPTOSIS
  • OXIDATION
  • RESONANCE
  • MAGNESIUM

Cite this

The covalent attachment of polyamines to proteins in plant mitochondria. / Votyakova, T V ; Wallace, H M ; Dunbar, B ; Wilson, S B .

In: European Journal of Biochemistry, Vol. 260, 1999, p. 250-257.

Research output: Contribution to journalArticle

Votyakova, T V ; Wallace, H M ; Dunbar, B ; Wilson, S B . / The covalent attachment of polyamines to proteins in plant mitochondria. In: European Journal of Biochemistry. 1999 ; Vol. 260. pp. 250-257.
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AU - Dunbar, B

AU - Wilson, S B

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N2 - Plant mitochondria from both potato end mung bean incorporated radioactivity into acid insoluble material when incubated with labelled polyamines (spermine, spermidine and putrescine). Extensive washing of mitochondrial precipitates with trichloroncetic acid and the excess of cold polyamine failed to remove bound radioactivity. Addition of nonradioactive polyamine stopped further incorporation of radioactivity but did net release radioactivity already bound. The radioactivity is incorporated into the membrane fraction, The labelling process has all the features of an enzymatic reaction: it is long lasting with distinctive kinetics peculiar to each polyamine, it is temperature dependent and is affected by N-ethylmaleimide, The latter inhibits the incorporation of putrescine but stimulates the incorporation of spermine and spermidine. Treatment of prelabelled mitochondria with pepsin releases bound radioactivity thus indicating protein to be the ligand for the attachment of polyamines. HPLC of mitochondrial hydrolysates revealed that the radioactivity bound to mitochondria is polyamines; traces of acetyl polyamines were also found in some samples. On autoradiograms of SDS/PAGE gels several radioactive bands of proteins were detected. Protein sequencing of labelled spots from a 2D gel gave a sequence which was 60% identical to catalase. We suggest that the attachment of polyamines to mitochondrial proteins occurs cotranslationally possibly via transglutaminases.

AB - Plant mitochondria from both potato end mung bean incorporated radioactivity into acid insoluble material when incubated with labelled polyamines (spermine, spermidine and putrescine). Extensive washing of mitochondrial precipitates with trichloroncetic acid and the excess of cold polyamine failed to remove bound radioactivity. Addition of nonradioactive polyamine stopped further incorporation of radioactivity but did net release radioactivity already bound. The radioactivity is incorporated into the membrane fraction, The labelling process has all the features of an enzymatic reaction: it is long lasting with distinctive kinetics peculiar to each polyamine, it is temperature dependent and is affected by N-ethylmaleimide, The latter inhibits the incorporation of putrescine but stimulates the incorporation of spermine and spermidine. Treatment of prelabelled mitochondria with pepsin releases bound radioactivity thus indicating protein to be the ligand for the attachment of polyamines. HPLC of mitochondrial hydrolysates revealed that the radioactivity bound to mitochondria is polyamines; traces of acetyl polyamines were also found in some samples. On autoradiograms of SDS/PAGE gels several radioactive bands of proteins were detected. Protein sequencing of labelled spots from a 2D gel gave a sequence which was 60% identical to catalase. We suggest that the attachment of polyamines to mitochondrial proteins occurs cotranslationally possibly via transglutaminases.

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KW - polyamines

KW - covalent modification

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KW - DERIVATIVES

KW - MEMBRANES

KW - APOPTOSIS

KW - OXIDATION

KW - RESONANCE

KW - MAGNESIUM

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JO - European Journal of Biochemistry

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