title: Evolution of substrate recognition sites (SRSs) in cytochromes P450 from Apiaceae exemplified by the CYP71AJ subfamily.
abstract: BACKGROUND
  Large proliferations of cytochrome P450 encoding genes resulting from gene duplications can be termed as 'blooms', providing genetic material for the genesis and evolution of biosynthetic pathways. Furanocoumarins are allelochemicals produced by many of the species in Apiaceaous plants belonging to the Apioideae subfamily of Apiaceae and have been described as being involved in the defence reaction against phytophageous insects.
  
  
  RESULTS
  A bloom in the cytochromes P450 CYP71AJ subfamily has been identified, showing at least 2 clades and 6 subclades within the CYP71AJ subfamily. Two of the subclades were functionally assigned to the biosynthesis of furanocoumarins. Six substrate recognition sites (SRS1-6) important for the enzymatic conversion were investigated in the described cytochromes P450 and display significant variability within the CYP71AJ subfamily. Homology models underline a significant modification of the accession to the iron atom, which might explain the difference of the substrate specificity between the cytochromes P450 restricted to furanocoumarins as substrates and the orphan CYP71AJ.
  
  
  CONCLUSIONS
  Two subclades functionally assigned to the biosynthesis of furanocoumarins and four other subclades were identified and shown to be part of two distinct clades within the CYP71AJ subfamily. The subclades show significant variability within their substrate recognition sites between the clades, suggesting different biochemical functions and providing insights into the evolution of cytochrome P450 'blooms' in response to environmental pressures.
authors.affiliation: University of Copenhagen, Department of Plant and Environmental Science, Copenhagen Plant Science Centre, Thorvaldsensvej 40, 1871, Frederiksberg C, Denmark. bjorn.dueholm@mymail.unisa.edu.au.|University of South Australia, School of Pharmacy and Medical Sciences, Adelaide, South Australia. bjorn.dueholm@mymail.unisa.edu.au.
authors.forename: Bjørn
authors.initials: B
authors.lastname: Dueholm
authors.affiliation: UMR1121 Université de Lorraine, 2 Avenue de la Forêt de Haye, 54518, Vandoeuvre-les-Nancy, France. celia.krieger@univ-lorraine.fr.|UMR1121 INRA, 2 Avenue de la Forêt de Haye, 54518, Vandoeuvre-les-Nancy, France. celia.krieger@univ-lorraine.fr.
authors.forename: Célia
authors.initials: C
authors.lastname: Krieger
authors.affiliation: University of Copenhagen, Department of Plant and Environmental Science, Copenhagen Plant Science Centre, Thorvaldsensvej 40, 1871, Frederiksberg C, Denmark. drew0072@uni.flinders.edu.au.
authors.forename: Damian
authors.initials: D
authors.lastname: Drew
authors.affiliation: UMR1121 Université de Lorraine, 2 Avenue de la Forêt de Haye, 54518, Vandoeuvre-les-Nancy, France. alexandre.olry@univ-lorraine.fr.|UMR1121 INRA, 2 Avenue de la Forêt de Haye, 54518, Vandoeuvre-les-Nancy, France. alexandre.olry@univ-lorraine.fr.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Olry
authors.affiliation: UMR1121 Université de Lorraine, 2 Avenue de la Forêt de Haye, 54518, Vandoeuvre-les-Nancy, France. tkamo@affrc.go.jp.|UMR1121 INRA, 2 Avenue de la Forêt de Haye, 54518, Vandoeuvre-les-Nancy, France. tkamo@affrc.go.jp.|National Institute for Agro-Environmental Sciences, 3-1-3 Kan-nondai, Tsukuba, 305-8604, Ibaraki, Japan. tkamo@affrc.go.jp.
authors.forename: Tsunashi
authors.initials: T
authors.lastname: Kamo
authors.affiliation: Technical University of Denmark, Centre for Biological and Sequence Analysis (CBS), Anker Engelunds Vej 1, 2800, Kgs. Lyngby, Denmark. olivier.taboureau@univ-paris-diderot.fr.|Molécules Thérapeutiques in silice (MTi), Inserm UMR-S 973 - Université Paris Diderot, Bat Lamarck A, 35 Rue Hélène Brion, 75205, Paris, France. olivier.taboureau@univ-paris-diderot.fr.
authors.forename: Olivier
authors.initials: O
authors.lastname: Taboureau
authors.affiliation: University of Copenhagen, Department of Plant and Environmental Science, Copenhagen Plant Science Centre, Thorvaldsensvej 40, 1871, Frederiksberg C, Denmark. corinna.weitzel@web.de.|UMR1121 Université de Lorraine, 2 Avenue de la Forêt de Haye, 54518, Vandoeuvre-les-Nancy, France. corinna.weitzel@web.de.|UMR1121 INRA, 2 Avenue de la Forêt de Haye, 54518, Vandoeuvre-les-Nancy, France. corinna.weitzel@web.de.
authors.forename: Corinna
authors.initials: C
authors.lastname: Weitzel
authors.affiliation: UMR1121 Université de Lorraine, 2 Avenue de la Forêt de Haye, 54518, Vandoeuvre-les-Nancy, France. frederic.bourgaud@univ-lorraine.fr.|UMR1121 INRA, 2 Avenue de la Forêt de Haye, 54518, Vandoeuvre-les-Nancy, France. frederic.bourgaud@univ-lorraine.fr.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Bourgaud
authors.affiliation: UMR1121 Université de Lorraine, 2 Avenue de la Forêt de Haye, 54518, Vandoeuvre-les-Nancy, France. alain.hehn@univ-lorraine.fr.|UMR1121 INRA, 2 Avenue de la Forêt de Haye, 54518, Vandoeuvre-les-Nancy, France. alain.hehn@univ-lorraine.fr.
authors.forename: Alain
authors.initials: A
authors.lastname: Hehn
authors.affiliation: University of Copenhagen, Department of Plant and Environmental Science, Copenhagen Plant Science Centre, Thorvaldsensvej 40, 1871, Frederiksberg C, Denmark. hts@plen.ku.dk.
authors.forename: Henrik Toft
authors.initials: HT
authors.lastname: Simonsen
mesh_terms: D000595:Amino Acid Sequence; D019661:Apiaceae / Q000737:chemistry / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000235:genetics*; D019143:Evolution, Molecular; D020440:Gene Duplication; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010802:Phylogeny; D016415:Sequence Alignment; D013379:Substrate Specificity
chemical_list: D003577:Cytochrome P-450 Enzyme System