Type I and type II fatty acid biosynthesis in Eimeria tenella: enoyl reductase activity and structure.
Apicomplexan parasites of the genus Eimeria are the major causative agent of avian coccidiosis, leading to high economic losses in the poultry industry. Recent results show that Eimeria tenella harbours an apicoplast organelle, and that a key biosynthetic enzyme, enoyl reductase, is located in this organelle. In related parasites, enoyl reductase is one component of a type II fatty acid synthase (FAS) and has proven to be an attractive target for antimicrobial compounds. We cloned and expressed the mature form of E. tenella enoyl reductase (EtENR) for biochemical and structural studies. Recombinant EtENR exhibits NADH-dependent enoyl reductase activity and is inhibited by triclosan with an IC50 value of 60 nm. The crystal structure of EtENR reveals overall similarity with other ENR enzymes; however, the active site of EtENR is unoccupied, a state rarely observed in other ENR structures. Furthermore, the position of the central beta-sheet appears to block NADH binding and would require significant movement to allow NADH binding, a feature not previously seen in the ENR family. We analysed the E. tenella genomic database for orthologues of well-characterized bacterial and apicomplexan FAS enzymes and identified 6 additional genes, suggesting that E. tenella contains a type II FAS capable of synthesizing saturated, but not unsaturated, fatty acids. Interestingly, we also identified sequences that appear to encode multifunctional type I FAS enzymes, a feature also observed in Toxoplasma gondii, highlighting the similarity between these apicomplexan parasites.
Lu
J Z
JZ
Department of Molecular Microbiology & Immunology, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD 21205, USA.
Muench
S P
SP
Allary
M
M
Campbell
S
S
Roberts
C W
CW
Mui
E
E
McLeod
R L
RL
Rice
D W
DW
Prigge
S T
ST
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D016786:Eimeria tenella / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D044943:Fatty Acid Desaturases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D054890:Fatty Acid Synthase, Type I / Q000378:metabolism*; D054889:Fatty Acid Synthase, Type II / Q000737:chemistry* / Q000378:metabolism*; D005227:Fatty Acids / Q000096:biosynthesis*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011994:Recombinant Proteins
D005227:Fatty Acids; D011994:Recombinant Proteins; D044943:Fatty Acid Desaturases; D054890:Fatty Acid Synthase, Type I; D054889:Fatty Acid Synthase, Type II
J Biol Chem. 1999 Apr 16;274(16):11110-4
10196195
Nature. 1999 Apr 1;398(6726):383-4
10201369
J Struct Biol. 1999 Apr-May;125(2-3):156-65
10222271
Biochemistry. 1999 Sep 21;38(38):12514-25
10493822
J Biol Chem. 1999 Oct 22;274(43):30811-7
10521472
Proc Natl Acad Sci U S A. 1999 Nov 9;96(23):13387-92
10557330
J Biol Chem. 1999 Dec 17;274(51):36465-71
10593943
Protein Sci. 1999 Nov;8(11):2529-32
10595560
Mol Biochem Parasitol. 2000 Feb 5;105(2):253-60
10693747
Microbiology. 2000 Feb;146 ( Pt 2):315-21
10708370
EMBO J. 2000 Apr 17;19(8):1794-802
10775264
Protein Expr Purif. 2000 Jul;19(2):312-8
10873547
J Biol Chem. 2001 Mar 2;276(9):6551-9
11050088
Nat Med. 2001 Feb;7(2):167-73
11175846
Proc Natl Acad Sci U S A. 2001 Feb 27;98(5):2723-8
11226307
J Parasitol. 2001 Feb;87(1):121-7
11227876
Mol Biol Evol. 2001 Mar;18(3):418-26
11230543
Int J Parasitol. 2001 Feb;31(2):109-13
11239932
J Biol Chem. 2001 Jul 27;276(30):28436-42
11319231
J Mol Biol. 2001 May 25;309(1):171-80
11491286
Annu Rev Microbiol. 2001;55:305-32
11544358
J Biol Chem. 2002 Apr 12;277(15):13106-14
11792710
J Mol Biol. 2002 May 31;319(2):257-74
12051904
Mol Biochem Parasitol. 2003 Feb;126(2):129-42
12615312
Lipids. 1976 Mar;11(3):209-15
1263764
Biochemistry. 2003 Apr 22;42(15):4406-13
12693936
J Med Chem. 2003 Apr 24;46(9):1627-35
12699381
Acta Crystallogr D Biol Crystallogr. 2003 Jul;59(Pt 7):1246-8
12832774
Proc Natl Acad Sci U S A. 2003 Nov 25;100(24):14281-6
14623959
Nat Rev Microbiol. 2004 Mar;2(3):203-16
15083156
Biochem J. 2004 Aug 1;381(Pt 3):719-24
15086316
J Comput Chem. 2004 Oct;25(13):1605-12
15264254
Acta Crystallogr D Biol Crystallogr. 1994 Sep 1;50(Pt 5):760-3
15299374
Mol Microbiol. 2005 Jan;55(1):39-53
15612915
Comb Chem High Throughput Screen. 2005 Feb;8(1):15-26
15720194
Eukaryot Cell. 2005 Apr;4(4):814-26
15821140
Annu Rev Biochem. 2005;74:791-831
15952903
Adv Parasitol. 2005;60:285-330
16230106
Cell Microbiol. 2006 Jan;8(1):1-9
16367861
Nature. 2006 May 18;441(7091):358-61
16710421
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 Jun 1;62(Pt 6):604-6
16754994
EMBO J. 2006 Jul 12;25(13):3214-22
16778769
Proc Natl Acad Sci U S A. 2006 Aug 29;103(35):13192-7
16920791
Int J Parasitol. 2007 Jan;37(1):33-51
17112527
Mol Microbiol. 2007 Mar;63(5):1331-44
17244193
Acta Crystallogr D Biol Crystallogr. 2007 Mar;63(Pt 3):328-38
17327670
Nat Rev Microbiol. 2007 Apr;5(4):287-97
17363967
Eukaryot Cell. 2007 Jun;6(6):984-96
17449654
J Mol Biol. 1990 Oct 5;215(3):403-10
2231712
Proc Natl Acad Sci U S A. 1989 Nov;86(21):8373-6
2682649
Methods Enzymol. 1997;276:307-26
27754618
FASEB J. 1994 Dec;8(15):1248-59
8001737
J Biol Chem. 1994 Feb 25;269(8):5493-6
8119879
Mol Gen Genet. 1994 Apr;243(2):249-52
8177222
Microbiol Rev. 1993 Sep;57(3):522-42
8246839
Structure. 1995 Sep 15;3(9):927-38
8535786
Trends Biochem Sci. 1995 Nov;20(11):478-80
8578593
Eur J Biochem. 1996 Dec 15;242(3):689-94
9022698
EMBO J. 1998 Mar 2;17(5):1183-91
9482715
Int J Parasitol. 1998 Jul;28(7):1089-98
9724880
Acta Crystallogr D Biol Crystallogr. 1998 Sep 1;54(Pt 5):905-21
9757107
Biochim Biophys Acta. 1998 Oct 2;1394(1):3-15
9767077
Proc Natl Acad Sci U S A. 1998 Oct 13;95(21):12352-7
9770490
Acta Pathol Microbiol Scand B. 1976 Dec;84B(6):401-13
998258