The covalent attachment of polyamines to proteins in plant mitochondria.
Plant mitochondria from both potato and mung bean incorporated radioactivity into acid insoluble material when incubated with labelled polyamines (spermine, spermidine and putrescine). Extensive washing of mitochondrial precipitates with trichloroacetic acid and the excess of cold polyamine failed to remove bound radioactivity. Addition of nonradioactive polyamine stopped further incorporation of radioactivity but did not 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.
Votyakova
T V
TV
Department of Molecular and Cell Biology, University of Aberdeen, Scotland.
Wallace
H M
HM
Dunbar
B
B
Wilson
S B
SB
D000595:Amino Acid Sequence; D002462:Cell Membrane / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000737:chemistry; D005033:Ethylmaleimide; D007887:Fabaceae / Q000737:chemistry; D007700:Kinetics; D008928:Mitochondria / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry*; D010946:Plants, Medicinal; D011073:Polyamines / Q000737:chemistry*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011700:Putrescine / Q000494:pharmacology; D011198:Solanum tuberosum / Q000737:chemistry; D013439:Sulfhydryl Reagents; D013696:Temperature
D010940:Plant Proteins; D011073:Polyamines; D011500:Protein Synthesis Inhibitors; D013439:Sulfhydryl Reagents; D010450:Endopeptidases; D005033:Ethylmaleimide; D011700:Putrescine