title: Comparative analysis of P450 signature motifs EXXR and CXG in the large and diverse kingdom of fungi: identification of evolutionarily conserved amino acid patterns characteristic of P450 family.
abstract: Cytochrome P450 monooxygenases (P450s) are heme-thiolate proteins distributed across the biological kingdoms. P450s are catalytically versatile and play key roles in organisms primary and secondary metabolism. Identification of P450s across the biological kingdoms depends largely on the identification of two P450 signature motifs, EXXR and CXG, in the protein sequence. Once a putative protein has been identified as P450, it will be assigned to a family and subfamily based on the criteria that P450s within a family share more than 40% homology and members of subfamilies share more than 55% homology. However, to date, no evidence has been presented that can distinguish members of a P450 family. Here, for the first time we report the identification of EXXR- and CXG-motifs-based amino acid patterns that are characteristic of the P450 family. Analysis of P450 signature motifs in the under-explored fungal P450s from four different phyla, ascomycota, basidiomycota, zygomycota and chytridiomycota, indicated that the EXXR motif is highly variable and the CXG motif is somewhat variable. The amino acids threonine and leucine are preferred as second and third amino acids in the EXXR motif and proline and glycine are preferred as second and third amino acids in the CXG motif in fungal P450s. Analysis of 67 P450 families from biological kingdoms such as plants, animals, bacteria and fungi showed conservation of a set of amino acid patterns characteristic of a particular P450 family in EXXR and CXG motifs. This suggests that during the divergence of P450 families from a common ancestor these amino acids patterns evolve and are retained in each P450 family as a signature of that family. The role of amino acid patterns characteristic of a P450 family in the structural and/or functional aspects of members of the P450 family is a topic for future research.
authors.affiliation: Department of Health Sciences, Faculty of Health and Environmental Sciences, Central University of Technology, Bloemfontein, Free State, South Africa.
authors.forename: Khajamohiddin
authors.initials: K
authors.lastname: Syed
authors.affiliation: Department of Health Sciences, Faculty of Health and Environmental Sciences, Central University of Technology, Bloemfontein, Free State, South Africa.
authors.forename: Samson Sitheni
authors.initials: SS
authors.lastname: Mashele
mesh_terms: D000595:Amino Acid Sequence; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000235:genetics*; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics; D005658:Fungi / Q000201:enzymology*
chemical_list: D005656:Fungal Proteins; D003577:Cytochrome P-450 Enzyme System
references.citation: Structure. 1995 Jan 15;3(1):41-62
references.pmid: 7743131
references.citation: BMC Genomics. 2012 Sep 02;13:444
references.pmid: 22937793
references.citation: Philos Trans R Soc Lond B Biol Sci. 2013 Jan 06;368(1612):20120428
references.pmid: 23297351
references.citation: Mol Biol Evol. 2011 Oct;28(10):2731-9
references.pmid: 21546353
references.citation: Genome Res. 2004 Jun;14(6):1188-90
references.pmid: 15173120
references.citation: Methods Mol Biol. 1998;107:15-24
references.pmid: 14577209
references.citation: Proc Natl Acad Sci U S A. 2001 May 8;98(10):5838-43
references.pmid: 11320210
references.citation: Biosci Biotechnol Biochem. 2007 Sep;71(9):2105-23
references.pmid: 17827683
references.citation: SAR QSAR Environ Res. 2001;12(4):359-66
references.pmid: 11769119
references.citation: J Biol Chem. 1997 Apr 11;272(15):9986-8
references.pmid: 9092539
references.citation: BMC Genomics. 2012 Oct 04;13:525
references.pmid: 23033934
references.citation: Antimicrob Agents Chemother. 2008 Oct;52(10):3718-24
references.pmid: 18663031
references.citation: Proc Natl Acad Sci U S A. 2012 Apr 3;109(14):5458-63
references.pmid: 22434909
references.citation: Hum Genomics. 2009 Oct;4(1):59-65
references.pmid: 19951895
references.citation: Xenobiotica. 1989 Oct;19(10):1149-60
references.pmid: 2683414
references.citation: Gene. 1989 Mar 15;76(1):121-36
references.pmid: 2663647
references.citation: Proteins. 2014 Mar;82(3):491-504
references.pmid: 24105833
references.citation: J Biol Chem. 1992 Jan 5;267(1):83-90
references.pmid: 1730627
references.citation: Science. 2012 Jun 29;336(6089):1715-9
references.pmid: 22745431
references.citation: Methods Mol Biol. 2006;320:1-10
references.pmid: 16719369
references.citation: Nucleic Acids Res. 2011 Jan;39(Database issue):D225-9
references.pmid: 21109532
references.citation: Nucleic Acids Res. 2004 Jul 1;32(Web Server issue):W327-31
references.pmid: 15215404
references.citation: Arch Microbiol. 2012 Apr;194(4):243-53
references.pmid: 21938516
references.citation: Yeast. 2000 Sep 15;16(12):1077-87
references.pmid: 10953079
references.citation: DNA Cell Biol. 1993 Jan-Feb;12(1):1-51
references.pmid: 7678494
references.citation: Mol Biol Evol. 1987 Nov;4(6):572-93
references.pmid: 3484338
references.citation: J Biochem. 1996 Mar;119(3):435-40
references.pmid: 8830036
references.citation: FEMS Microbiol Lett. 2003 Apr 11;221(1):103-10
references.pmid: 12694917
references.citation: FEMS Microbiol Lett. 2001 Sep 11;203(1):11-21
references.pmid: 11557134
references.citation: Crit Rev Microbiol. 2012 Nov;38(4):339-63
references.pmid: 22624627
references.citation: FEBS Lett. 2006 Nov 27;580(27):6338-42
references.pmid: 17092500
references.citation: J Biol Chem. 1991 Feb 25;266(6):3372-5
references.pmid: 1899862
references.citation: Annu Rev Biochem. 1987;56:945-93
references.pmid: 3304150
references.citation: PLoS One. 2014 Jan 22;9(1):e86683
references.pmid: 24466198
references.citation: Biochem Biophys Res Commun. 2010 Sep 3;399(4):492-7
references.pmid: 20674550
references.citation: Philos Trans R Soc Lond B Biol Sci. 2013 Jan 06;368(1612):20120434
references.pmid: 23297356
references.citation: FEBS Lett. 1996 Jul 8;389(3):268-72
references.pmid: 8766713
references.citation: Nucleic Acids Res. 2009 Jan;37(Database issue):D205-10
references.pmid: 18984618
references.citation: BMC Genomics. 2013 Feb 27;14:130
references.pmid: 23442312
references.citation: J Biotechnol. 2006 Jun 25;124(1):128-45
references.pmid: 16516322
references.citation: BMC Genomics. 2008 Aug 28;9:402
references.pmid: 18755027
references.citation: J Agric Food Chem. 2006 Dec 13;54(25):9398-404
references.pmid: 17147424
references.citation: Nat Commun. 2012 Jun 26;3:913
references.pmid: 22735441
references.citation: PLoS One. 2011;6(12):e28286
references.pmid: 22164262
references.citation: Biochim Biophys Acta. 2007 Mar;1770(3):467-77
references.pmid: 16963187
references.citation: Mol Cell Endocrinol. 2004 Feb 27;215(1-2):165-70
references.pmid: 15026190
references.citation: Appl Environ Microbiol. 2005 Jun;71(6):3192-8
references.pmid: 15933021
references.citation: Appl Environ Microbiol. 2013 Apr;79(8):2692-702
references.pmid: 23416995
references.citation: Nucleic Acids Res. 1990 Oct 25;18(20):6097-100
references.pmid: 2172928
references.citation: Appl Environ Microbiol. 2003 Oct;69(10):5992-9
references.pmid: 14532054
references.citation: Mycologia. 2013 Nov-Dec;105(6):1445-55
references.pmid: 23928414
references.citation: BMC Struct Biol. 2010 Oct 15;10:34
references.pmid: 20950472
references.citation: Biochem Biophys Res Commun. 2011 Apr 1;407(1):118-23
references.pmid: 21362401
references.citation: Biochim Biophys Acta. 2011 Jan;1814(1):14-8
references.pmid: 20736090
references.citation: Philos Trans R Soc Lond B Biol Sci. 2013 Jan 06;368(1612):20120430
references.pmid: 23297353
references.citation: DNA. 1987 Feb;6(1):1-11
references.pmid: 3829886
references.citation: Arch Biochem Biophys. 1999 Sep 1;369(1):24-9
references.pmid: 10462437
references.citation: Bioinformatics. 2007 Aug 1;23(15):2015-7
references.pmid: 17510166