Molecular and biochemical characterization of tomato farnesyl-protein transferase.
The prenylation of membrane-associated proteins involved in the regulation of eukaryotic cell growth and signal transduction is critically important for their subcellular localization and biological activity. In contrast to mammalian cells and yeast, however, the function of protein prenylation in plants is not well understood and only a few prenylated proteins have been identified. We partially purified and characterized farnesyl-protein transferase from tomato (Lycopersicon esculentum, LeFTase) to analyze its biochemical and molecular properties. Using Ras- and G gamma-specific peptide substrates and competition assays we showed that tomato protein extracts have both farnesyl-protein transferase and geranylgeranyl-protein transferase 1 activities. Compared with the heterologous synthetic peptide substrates, the plant-specific CaaX sequence of the ANJ1 protein is a less efficient substrate for LeFTase in vitro. LeFTase activity profiles and LeFTase beta-subunit protein (LeFTB) levels differ significantly in various tissues and are regulated during fruit development. Partially purified LeFTase requires Zn2+ and Mg2+ for enzymatic activity and has an apparent molecular mass of 100 kD Immunoprecipitation experiments using anti-alpha LeFTB antibodies confirmed that LeFTB is a component of LeFTase but not of tomato geranylgeranyl-protein transferase 1. Based on their conserved bio-chemical activities, we expect that prenyltransferases are likely integrated with the sterol biosynthesis pathway in the control of plant cell growth.
Schmitt
D
D
Department of Plant Biology, University of California, Berkeley 94720, USA.
Callan
K
K
Gruissem
W
W
D019883:Alkyl and Aryl Transferases; D019204:GTP-Binding Proteins / Q000378:metabolism; D007527:Isoenzymes / Q000378:metabolism; D018551:Solanum lycopersicum / Q000201:enzymology* / Q000254:growth & development; D009842:Oligopeptides / Q000378:metabolism*; D010940:Plant Proteins / Q000378:metabolism; D017368:Protein Prenylation; D013379:Substrate Specificity; D014018:Tissue Distribution; D014166:Transferases / Q000187:drug effects / Q000378:metabolism*; D015032:Zinc / Q000494:pharmacology; D018631:ras Proteins / Q000378:metabolism
C083280:ANJ1 protein, Atriplex nummularia; D007527:Isoenzymes; D009842:Oligopeptides; D010940:Plant Proteins; D014166:Transferases; D019883:Alkyl and Aryl Transferases; C083291:geranylgeranyltransferase type-I; C064911:p21(ras) farnesyl-protein transferase; D019204:GTP-Binding Proteins; D018631:ras Proteins; D015032:Zinc