Target+-+Enoyl-acyl-carrier-protein+reductase+(Plasmodium+falcifarum+3D7)

 ***NCBI Gene # or RefSeq#:**46361102  ***Protein ID (NP or XP #) or Wolbachia#:**XP_966137.1
 * *Target (protein/gene name): **** enoyl-acyl carrier reductase **
 * Organism (including strain):** **//Plasmodium falciparum 3D7//**
 * Etiologic Risk Group (see link below):** **Risk group level 3**
 * [] **

//Plasmodium falciparum// is a protozoan parasite that causes the most dangerous strain of malaria in humans. In 2012, malaria caused about 627,000 deaths mostly among African children. This disease is transmitted only through Anopheles mosquito bites and is most prevalent in tropical areas where the mosquitoes can breed in shallow water. Malaria is curable and preventable and recent studies have shown that the enoyl reductase enzyme (ENR) that is responsible for type II fatty-acid biosynthesis pathway in the protozoa has been inhibited by the antibacterial agent Triclosan.
 * *Background/ ****Disease Information **** (sort of like the Intro to your Mini Research Write up): **


 * Link to TDR Targets page** **(if present):** http://tdrtargets.org/targets/view?gene_id=4502
 * Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.)** http://plasmodb.org/plasmo/showRecord.doname=GeneRecordClasses.GeneRecordClass&primary_key=PFF0730c

PFF0730c has essentiality data Gene/Ortholog: mtu1508 (OG4_13747); Phenotype: non-essential; Source study: nmpdr Gene/Ortholog: eco1251 (OG4_13747); Phenotype: undefined; Source study: blattner Gene/Ortholog: eco1251 (OG4_13747); Phenotype: essential; Source study: gerdes Gene/Ortholog: eco1251 (OG4_13747); Phenotype: essential; Source study: keio Gene/Ortholog: eco1251 (OG4_13747); Phenotype: essential; Source study: shigen
 * Essentiality of this protein:**

The enoyl-acyl carrier protein reductase catalyzes the final step in fatty acid elongation type II pathway in Plasmodium falciparum. Source: []


 * Complex of proteins?: ** in complex with NADH
 * Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): ****0.8 **

 **Link to BRENDA EC# page: ****[] **  **-- ** Show screenshot of BRENDA enzyme mechanism schematic
 * *EC#: ** **1.3.1.9 **

-- link to Sigma (or other company) page for assay (see Sigma links below) http://www.sigmaaldrich.com/catalog/product/sigma/28007?lang=en®ion=US -- links to assay reagents (substrates) pages. http://www.brenda-enzymes.info/literature/lit.php4?e=1.3.1.9&r=390898 --- List cost and quantity of substrate reagents, supplier, and catalog #
 * Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):**
 * Substrate reagent**: Crotonoyl coenzyme A trilithium salt
 * Cost:** 5mg = $266.50 ; 25mg = $920.00
 * Supplier**: Sigma-Aldrich
 * Catalog #**: MFCD00151226

 -- PDB # or closest PDB entry if using homology model: 1NHW
 * Structure Available (PDB or Homology model) **

<span style="font-family: 'Times New Roman',serif; font-size: 11pt;"> **<span style="background-color: white; background-position: initial initial; background-repeat: initial initial; box-sizing: border-box;">Expression Information (has it been expressed in bacterial cells): **
 * <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt;">Current Inhibitors: **<span style="background-color: #f5f5f5; background-position: initial initial; background-repeat: initial initial; font-family: 'Times New Roman',serif; font-size: 11pt;">5-chloro-2-(2,4-dichlorophenoxy)phenol
 * Although the TDR page showed no information on expression, BRENDA shows that this protein has been expressed in //Escherichia coli// **



There are two different vectors that can be used to purify the protein: pMALc2x and pMALcHT vectors. __pMALc2x__: Cells were resuspended in lysis buffer (20mM sodium/potassium phosphate pH 7.5, 1 mg/mL lysozyme, 2.5 micrograms/mL DNAse I, 200 mM NaCl), centrifuged, and applied to a 10mL amylose column for affinity purification. Purified MBP-ENR fusion protein was digested with factor Xa and desalted with a HiPrep 26/10 desalting column and applied to an SP Sepharose cation-exchange column. __pMALcHT:__ Cells were clarified by centrifugation and applied to a 5 mL HiTrap Chelating HP colum and desalted with a HiPrep 26/10 desalting column and loaded onto a 5mL HiTrap SP Fast Flow column. Source: http://journals.iucr.org/d/issues/2003/07/00/za0101/za0101.pdf
 * Purification Method:**


 * <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt;">Image of protein (PyMol with features delineated and shown separately): **

MNKISQRLLFLFLHFYTIVCFIQNNTQKTFHNVLQNEQIRGKEKAFYRKEKRENIFIGNK MKHLNNMNNTHNNNHYMEKEEQDASNINKIKEENKNEDICFIAGIGDTNGYGWGIAKELSKRNVKIIFGIWPPVYNIFMKNYKNGKFDNDMIIDKDKKMNILDMLPFDASFDTANDIDEETKNNKRYNMLQNYTIEDVANLIHQKYGKINMLVHSLANAKEVQKDLLNTSRKGYLDALSKSSYSLISLCKYFVNIMKPQSSIISLTYHASQKVVPGYGGGMSSAKAALESDTRVLAYHLGRNYNIRINTISAGPLKSRAATAINKLNNTYENNTNQNKNRNSHDVHNIMNNSGEKEEKKNSASQNYTFIDYAIEYSEKYAPLRQKLLSTDIGSVASFLLSRESRAITGQTIYVDNGLNIMFLPDDIYRNENE
 * <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt;">*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): **

**<span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt;">*length of your protein in Amino Acids: ****<span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; font-weight: normal;">432 amino acids ** **<span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-family: 'Times New Roman',serif; font-size: 11pt;">Molecular Weight ** **<span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt;">of your protein in kiloDaltons using the ****<span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt;">[|Expasy ProtParam] ****<span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt;">website ****<span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; font-weight: normal;">49.7623 kDA ** Ext. coefficient 45395 <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt;">Abs 0.1% (=1 g/l) 0.912, assuming all pairs of Cys residues form cystines
 * <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt;">Molar ** **<span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-family: 'Times New Roman',serif; font-size: 11pt;">Extinction coefficient ** **<span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt;">of your protein at 280 nm wavelength: **

<span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt;">Ext. coefficient 45270 <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt;">Abs 0.1% (=1 g/l) 0.910, assuming all Cys residues are reduced

<span style="background: white; tab-stops: 45.8pt 91.6pt 137.4pt 183.2pt 229.0pt 274.8pt 320.6pt 366.4pt 412.2pt 458.0pt 503.8pt 549.6pt 595.4pt 641.2pt 687.0pt 732.8pt;">** TMpred graph Image ** <span style="font-family: 'Times New Roman',serif; font-size: 11pt; line-height: 12.65pt;">(@http://www.ch.embnet.org/software/TMPRED_form.html) <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; line-height: 0px; overflow: hidden;">

1 atgaataaaa tatcacaacg gttattattc ctctttctac atttttatac cattgtgtgc <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; line-height: 12.65pt;"> 61 tttatccaga ataatactca aaagacattc cataacgttc ttcagaatga acagatacga <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; line-height: 12.65pt;"> 121 ggtaaggaaa aagcatttta taggaaagaa aaacgcgaaa acatatttat aggaaacaaa <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; line-height: 12.65pt;"> 181 atgaaacact taaataatat gaataataca cacaataata atcattatat ggagaaagaa <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; line-height: 12.65pt;"> 241 gaacaagatg catcaaacat aaacaaaatt aaagaagaaa ataaaaatga agatatttgt <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; line-height: 12.65pt;"> 301 tttattgctg gtataggaga tacaaatgga tacgggtggg gtattgctaa agaattatcc <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; line-height: 12.65pt;"> 361 aaaagaaatg taaaaataat tttcggtatt tggcctcctg tttataatat ttttatgaaa <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; line-height: 12.65pt;"> 421 aattataaaa acgggaaatt tgataatgat atgattattg ataaagataa gaaaatgaat <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; line-height: 12.65pt;"> 481 atattagaca tgctaccctt tgacgcttct tttgatactg ccaatgatat agatgaagaa <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; line-height: 12.65pt;"> 541 acgaaaaata ataaaagata taatatgtta cagaattata ctatagaaga tgttgccaat <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; line-height: 12.65pt;"> 601 ttaatacatc aaaaatatgg aaaaattaat atgctcgttc attctttagc taacgctaaa <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; line-height: 12.65pt;"> 661 gaagttcaaa aagatttatt aaataccagt cgaaaaggtt atttagatgc tcttagtaaa <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; line-height: 12.65pt;"> 721 agttcttact ctttaatatc attatgtaaa tattttgtca atattatgaa acctcaatcc <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; line-height: 12.65pt;"> 781 agcattattt cattaacata tcatgctagc caaaaagttg tgccaggcta tggtggaggt <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; line-height: 12.65pt;"> 841 atgtccagcg ctaaagctgc actcgaatct gataccagag ttttagctta tcatctagga <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; line-height: 12.65pt;"> 901 agaaattata atatcagaat taatactatc agtgcaggac cacttaaatc aagagcagct <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; line-height: 12.65pt;"> 961 actgctatta ataaattaaa caatacatat gaaaataata caaatcaaaa taaaaatagg <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; line-height: 12.65pt;"> 1021 aatagtcatg atgttcataa tattatgaac aattcaggtg aaaaggaaga aaaaaaaaat <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; line-height: 12.65pt;"> 1081 tcagctagcc aaaattatac atttatagat tatgcaatag agtattcaga aaagtatgct <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; line-height: 12.65pt;"> 1141 ccattacgtc aaaagttatt atcaacagat attggatctg tagcatcatt tttgttatct <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; line-height: 12.65pt;"> 1201 cgagagagtc gtgcgataac tgggcagaca atatatgtgg ataacggatt aaatataatg <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt; line-height: 12.65pt;"> 1261 tttttaccag atgatatata tcgcaatgaa aatgaataa
 * CDS gene sequence (paste as text only):**

<span style="font-family: 'Times New Roman',serif; font-size: 11pt;"> ** *CDS Gene Sequence (codon optimized) - copy from ** ** output ** ** of ** ** Primer Design ** ** Protocol (paste as text only): N/A **
 * <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-size: 11pt;">*GC% Content for gene: **<span style="font-family: 'Times New Roman',serif; font-size: 11pt;"> **<span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-weight: normal;">28.098537336413% **
 * *GC% Content for gene (codon optimized): N/A **

<span style="background-color: yellow; background-position: initial initial; background-repeat: initial initial; font-family: 'Times New Roman',serif; font-size: 11pt;">Do Not Need this info for Spring (but still copy these lines to your Target page for now) <span style="font-family: 'Times New Roman',serif; font-size: 11pt;"> ** Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): ** <span style="font-family: 'Times New Roman',serif; font-size: 11pt;"> ** ( ** **<span style="background-color: white; background-position: initial initial; background-repeat: initial initial; color: #008000; font-size: 11pt;">link to DNA Works output ** **<span style="background-color: white; background-position: initial initial; background-repeat: initial initial; color: #008000; font-family: 'Times New Roman',serif; font-size: 11pt;">text file ** **<span style="background-color: white; background-position: initial initial; background-repeat: initial initial; color: #008000; font-size: 11pt;">- **<span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-family: 'Times New Roman',serif; font-size: 11pt;">that should be saved in your Google Docs folder after you did the primer design protocol) <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-family: 'Times New Roman',serif; font-size: 11pt;"> -- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.

<span style="font-family: 'Times New Roman',serif; font-size: 11pt;"> ** Primer design results for 'tail' primers (this is just 2 sequences): ** <span style="background-color: white; background-position: initial initial; background-repeat: initial initial; font-family: 'Times New Roman',serif; font-size: 11pt;"> ** code

code

code

code