Purification, Characterization and Partial Amino Acid Sequencing of Hydroxycinnamoyl-CoA: Tyramine N-(hydroxycinnamoyl)transferase from Tobacco Cell-Suspension Cultures (original) (raw)
Related papers
Biochimica et biophysica acta, 1996
The cleavage of purified bovine alpha s1-casein separately by cardosin A and cardosin B, two distinct milk-clotting aspartic proteinases (APs) present in the stigmas of the plant Cynara cardunculus L., was studied. Casein digestion peptides were separated either by SDS-PAGE or by reverse-phase HPLC, and their N-terminal amino acid sequences were subsequently determined by automated Edman degradation, thus identifying the cleavage sites. Results showed that both enzymes exert a similar but distinct action on bovine alpha s1-casein. In common they have the preference for the bond Phe23-Phe24, and the cleavage of Trp164-Tyr165 and Phe153-Tyr154. Cardosin A also cleaves the bond Tyr165-Tyr166, whereas Cardosin B cleaves an extra type of bond, Phe150-Arg151, revealing a slightly broader specificity. A model for the action of both enzymes on bovine alpha s1-casein is proposed and discussed. In comparison with the reported action of chymosin on bovine alpha s1-casein, both cardosins proved...
Protoplasma, 2008
Following on from previous work, the temporal and spatial accumulation of the aspartic proteinases (EC 3.4.23) cardosin A and cardosin B during postembryonic seed development of cardoon (Cynara cardunculus) was studied. mRNA and protein analyses of both cardosins suggested that the proteins accumulate during seed maturation, and that cardosin A is later synthesised de novo at the time of radicle emergence. Immunocytochemistry revealed that the precursor form of cardosin A accumulates in protein bodies and cell walls. This localisation in seeds is different from that previously described for cardoon flowers, suggesting a tissue-dependent targeting of the protein. It is known that procardosins are active and may have a role in proteolysis and processing of storage proteins. However, the presence of procardosin A in seeds could be related to the proposed role of the plant-specific insert in membrane lipid conversion during water uptake and solute leakage in actively growing tissues. This is in accordance with the recently proposed bifunctional role of aspartic proteinase precursor molecules that possess a membrane-destabilising domain in addition to a protease domain. Mature cardosin B, but not its mRNA, was detected in the first hours after seed imbibition and disappeared at the time of radicle emergence. This extracellular aspartic protease has already been implicated in cell wall loosening and remodelling, and its role in seed germination could be related to loosening tissue constraints for radicle protusion. The described pattern of cardosin A and B expression suggests a finely tuned developmental regulation and prompts an analysis of their possible roles in the physiology of postembryonic development.