title: Subcellular location of the tetrapyrrole synthesis enzyme porphobilinogen deaminase in higher plants: an immunological investigation.
abstract: A recombinant plasmid, pArab8, harbouring the cDNA encoding the mature form of the tetrapyrrole synthesis enzyme porphobilinogen deaminase (EC 4.3.1.8; also known as hydroxymethylbilane synthase) from Arabidopsis thaliana (L.) Heynh. has been constructed, and used to transform Escherichia coli. The porphobilinogen deaminase protein from Arabidopsis was overexpressed in this strain, and purified to homogeneity (3000-fold) with a yield of 20%. Antibodies were raised against the purified plant enzyme, and used in Western blot analysis, immunoprecipitation of enzyme activity and immuno-gold electron microscopy. The results indicate that the enzyme is confined to plastids in both leaves and roots. The implications of this finding for plant tetrapyrrole synthesis are discussed.
authors.affiliation: Department of Plant Sciences, University of Cambridge, UK.
authors.forename: M
authors.initials: M
authors.lastname: Witty
authors.forename: R M
authors.initials: RM
authors.lastname: Jones
authors.forename: M S
authors.initials: MS
authors.lastname: Robb
authors.forename: P M
authors.initials: PM
authors.lastname: Jordan
authors.forename: A G
authors.initials: AG
authors.lastname: Smith
mesh_terms: D000818:Animals; D017360:Arabidopsis / Q000378:metabolism; D015153:Blotting, Western; D003001:Cloning, Molecular; D004926:Escherichia coli; D011163:Hydroxymethylbilane Synthase / Q000235:genetics / Q000378:metabolism*; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism
chemical_list: D011993:Recombinant Fusion Proteins; D011163:Hydroxymethylbilane Synthase