Investigations about N-aminopropyl transferases probably involved in biomineralization.
Polyamines are widespread distributed all over in living organisms. In Thalassiosira pseudonana 10 N-aminopropyl transferase like nucleotide sequences exists. It is assumed that these sequences are involved in the biomineralization of the diatom shell. The cDNA of the sequences were cloned, recombinant overexpressed and assayed with decarboxylated S-adenosylmethionine and several radioactive labelled polyamines. However, only a spermidine synthase and a thermospermine synthase were found to be enzymatically active in an in vitro assay. Both enzyme activities could be recognized in the crude extracts of Thalassiosira pseudonana and Cyclotella meneghiana. In further investigations the kinetics of the thermospermine synthase was determined and a site-specific mutagenesis of the bindig cavity of decarboxylated S-adenosylmethionine was carried out.
Romer
P
P
Department of Orthodontics, Regensburg University, Germany. peter.proff@klinik.uni-regensburg.de
Faltermeier
A
A
Mertins
V
V
Gedrange
T
T
Mai
R
R
Proff
P
P
D000595:Amino Acid Sequence; D015317:Biogenic Polyamines / Q000096:biosynthesis*; D002855:Chromatography, Thin Layer; D003001:Cloning, Molecular; D017377:Diatoms / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction; D011436:Spermidine Synthase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D013097:Spermine Synthase / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic
D015317:Biogenic Polyamines; D011436:Spermidine Synthase; D013097:Spermine Synthase