A novel C-S lyase from the latex-producing plant Taraxacum brevicorniculatum displays alanine aminotransferase and l-cystine lyase activity.
We isolated a novel pyridoxal-5-phosphate-dependent l-cystine lyase from the dandelion Taraxacum brevicorniculatum. Real time qPCR analysis showed that C-S lyase from Taraxacum brevicorniculatum (TbCSL) mRNA is expressed in all plant tissues, although at relatively low levels in the latex and pedicel. The 1251 bp TbCSL cDNA encodes a protein with a calculated molecular mass of 46,127 kDa. It is homologous to tyrosine and alanine aminotransferases (AlaATs) as well as to an Arabidopsis thaliana carbon-sulfur lyase (C-S lyase) (SUR1), which has a role in glucosinolate metabolism. TbCSL displayed in vitrol-cystine lyase and AlaAT activities of 4 and 19nkatmg(-1) protein, respectively. However, we detected no in vitro tyrosine aminotransferase (TyrAT) activity and RNAi knockdown of the enzyme had no effect on phenotype, showing that TbCSL substrates might be channeled into redundant pathways. TbCSL is in vivo localized in the cytosol and functions as a C-S lyase or an aminotransferase in planta, but the purified enzyme converts at least two substrates specifically, and can thus be utilized for further in vitro applications.
Institut für Biologie und Biotechnologie der Pflanzen, Westfälische Wilhelms-Universität Münster, Hindenburgplatz 55, 48143 Münster, Germany.
Oliver
O
Munt
Dirk
D
Prüfer
Christian
C
Schulze Gronover
D000410:Alanine Transaminase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000595:Amino Acid Sequence; D053898:Biosynthetic Pathways; D003600:Cytosol / Q000201:enzymology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D035264:Flowers / Q000166:cytology / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D015870:Gene Expression; D018506:Gene Expression Regulation, Plant / Q000235:genetics*; D055785:Gene Knockdown Techniques; D007840:Latex / Q000378:metabolism; D008190:Lyases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D018515:Plant Leaves / Q000166:cytology / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D018517:Plant Roots / Q000166:cytology / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D030821:Plants, Genetically Modified; D011993:Recombinant Fusion Proteins; D036226:Seedlings / Q000166:cytology / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D016415:Sequence Alignment; D013379:Substrate Specificity; D031205:Taraxacum / Q000166:cytology / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism
D007840:Latex; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D000410:Alanine Transaminase; D008190:Lyases; C033511:carbon-sulfur lyase