ABSTRACT
The four-step caffeine biosynthetic pathway includes three methylation steps that utilise S-adenosyl-L-methionine (SAM) as the methyl donor. In the process SAM is converted to S-adenosyl-L-homocysteine (SAH) which in turn is hydrolysed to L-homocysteine and adenosine. Significant amounts of radioactivity from [methyl-(14)C]methionine and [methyl-(14)C]SAM were incorporated into theobromine and caffeine in young tea leaf segments, and very high SAH hydrolase activity was found in cell-free extracts from young tea leaves. Substantial amounts of radioactivity from [adenosyl-(14)C]SAH were also recovered as theobromine and caffeine in tea leaf segments, indicating that adenosine derived from SAH is utilised for the synthesis of the purine ring of caffeine. From the profiles of activity of related enzymes in tea leaf extracts, it is proposed that the major route from SAM to caffeine is a SAM-->SAH-->adenosine-->adenine-->AMP-->IMP-->XMP-->xanthosine-->7-methylxanthosine-->7-methylxanthine-->theobromine-->caffeine pathway. In addition, direct adenosine kinase-catalysed formation of AMP from adenosine may participate as an alternative minor route. The activity of two of the three N-methyltransferase activities involved in caffeine biosynthesis and part of the activities of SAH hydrolase, adenosine nucleosidase, adenine phosphoribosyltransferase and adenosine kinase were located in tea chloroplasts. In contrast, no detectable activity of SAM synthetase was associated with the purified chloroplast fraction. This is a first demonstration that the purine skeleton of caffeine is synthesised from adenosine released from the SAM cycle.
Subject(s)
Adenosine/metabolism , Caffeine/metabolism , Plant Leaves/metabolism , S-Adenosylmethionine/metabolism , Tea/metabolism , Adenosylhomocysteinase , Caffeine/chemistry , Carbon Dioxide/metabolism , Cell Extracts , Chloroplasts/enzymology , Chloroplasts/metabolism , Homocysteine/metabolism , Hydrolases/metabolism , Methionine Adenosyltransferase/metabolism , Methylation , Plant Leaves/cytology , Plant Leaves/enzymology , Plant Leaves/growth & development , S-Adenosylhomocysteine/metabolism , Tea/cytology , Tea/enzymology , Tea/growth & development , Theobromine/metabolismABSTRACT
Cytological changes in the cementoblasts of rats treated with vincristine were investigated. Accumulation of secretion granules indicated a disturbed secretory mechanism in the cells. Conspicuous dilation of the endoplasmic reticulum suggested disturbance in the transport of substances from the rough endoplasmic reticulum to the Golgi elements. These disturbances were interpreted to be a result of disruption of microtubules.
