Target+Sp15+-+Adenylosccinate+synthetase+(P.+falciparum)

** Disease Information ** ** (sort of like the Intro to your Mini **** Research Write **** up: ** Malaria is caused by a parasite //Plasmodium falciparum//. The protozoan parasite is infects the humans thru a vector of mostly mosquitos. Malaria symptoms include fever, chills, and flu-like illness. When untreated the disease can worsen leading to complication and even death. []  []
 * Target (protein/gene name): ** adenylosuccinate synthetase
 * *NCBI Gene # or RefSeq#: ** NCBI Gene ID:814251
 * *Protein ID (NP or XP #) or Wolbachia#: ** XP_001350257.1
 * *Organism (including strain): ** P. falciparum 3D7
 * Etiologic Risk Group (see link below ): ** India, Africa, and Some Areas in South Americ ** a ** and is often seen in pregnant women, children under 5 years, people with HIV/AIDS and non-immune travelers.
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 * Link to TDR Targets page (if present): ** []
 * Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.) **

***EC#:** 6.3.4.4
 * Essentiality of this protein: **
 * ** “ ** catalysing the first committed step in the synthesis of AMP from IMP”
 * “in regulation of protein function” [|http://www.sciencedirect.com/science/article/pii/S0022283603014426#]
 * Is it a monomer or multimer as biological unit **** ? (make prediction at **** http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html ): ** Monomer
 * Complex of proteins: ** Simple only one chain
 * Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): ** 0.8 from TDR Targets
 * Link to BRENDA EC# page: [|http://www.brenda-enzymes.org/enzyme.php?ecno=6.3.4.4#] **
 * NATURAL SUBSTRATES ** [[image:Screen Shot 2015-04-18 at 2.55.38 PM.png width="800" height="213"]]

** Enzyme Assay ** ** information (spectrophotometric, coupled assay ?, reagents): ** Spectrophotometric **-- link (or citation) to paper that contains assay information:** “AMP deaminase (EC 3.5.4.6) [33] ; adenylosuccinate synthetase (EC 6.3.4.4) [34] ; adenylosuccinate lyase (EC 4.3.2.2) [35] ; IMP dehydrogenase (EC 1.2.1.14)” [] **--****links to assay reagents (substrates) pages.**[] **---** Li st cost and quantity of substrate reagents, supplier, and catalog #: HEPES Buffer: BP299-100 on Fisher $83.11 for 100mL

IMP: AC22626-0050 on Fisher 28.26 for 5g

GTP: ICN10071010 on Fisher 32.16 for 10g

MgCl2 : BP6114 on Fisher for $14.70 for 1mL, 25mM

**Structure (PDB or Homology model)** -- PDB # or closest PDB entry if using homology model: 1P9B
 * Current Inhibitors **** : Th ** e presence, or over abundance of AMP, GMP, GDP and adenylosuccinate acts as a feedback inhibitor http://www.proteopedia.org/wiki/index.php/Adenylosuccinate_Synthetase

** Purification Method ** ** : ** Ornstein and Davis (16), with 7% polyacrylamidegels, pH 8.3 Tris-glycine buffer and 20 to 100 pg of protein.
 * Expression Information (has it been expressed in bacterial cells): ** partially purified with //E. coli-// homogeneous preparation of the E. coli enzyme using ammonium sulfate fractionation and DEAE-cellulose and Sephadex G-100 chromatography.
 * [] **

MAIFDHQIKN VDKGN VVAIL G AQWG D EG KG KIIDMLSEY S D ITCR F N GG A N AG HTISV ND KKYAL HLL P C GVLYDN N ISV L G NG M VI H VK SLMEEIESV G  GK LLDR LYLS NKA HI LF DIH QIIDSIQETK KLK EG KQIGT T KRG IGPCYS TKA SR IGI R L GTLK N FENFK NMYSKLIDHL MDL YN ITEYD KEKELNLFYN YHIKLRDR IV D VISFMNTNL E NN KK VLIE G A N A AML D IDF GTYPYV T SS C T T VGGVFS GL G IH HKK LN LV VGVVK SY LTR V GCG PF LT E L N N DVGQYLRE K GH E Y G TTT K R P R RCG W LD I PMLLYVKCIN SI D MINLT K L DVL SG L E EIL LCVNFKN KKT GE L LE KG CYP V E EEIS EE YE PVYEKFS GWK E DI ST CN E FD E L P ENAKKYI LAIEKYL K TP IVWIG V GPNR K N MIV KKNFN LN
 * Image of protein (PyMol with features delineated and shown separately): **
 * Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): **

** Molecular Weight ** ** of your protein in kiloDaltons using the ** [|**Expasy ProtParam**]**website:** 50.0218 kDA
 * *length of your protein in Amino Acids: ** 442 Amino Acids
 * Molar ** ** Extinction coefficient ** ** of your protein at 280 nm wavelength: ** 1.078

atggcgatttttgatcatcagattaaaaacgtggataaaggcaacgtggtggcgattctg ggcgcgcagtggggcgatgaaggcaaaggcaaaattattgatatgctgagcgaatatagc gatattacctgccgctttaacggcggcgcgaacgcgggccataccattagcgtgaacgat aaaaaatatgcgctgcatctgctgccgtgcggcgtgctgtatgataacaacattagcgtg ctgggcaacggcatggtgattcatgtgaaaagcctgatggaagaaattgaaagcgtgggc ggcaaactgctggatcgcctgtatctgagcaacaaagcgcatattctgtttgatattcat cagattattgatagcattcaggaaaccaaaaaactgaaagaaggcaaacagattggcacc accaaacgcggcattggcccgtgctatagcaccaaagcgagccgcattggcattcgcctg ggcaccctgaaaaactttgaaaactttaaaaacatgtatagcaaactgattgatcatctg atggatctgtataacattaccgaatatgataaagaaaaagaactgaacctgttttataac tatcatattaaactgcgcgatcgcattgtggatgtgattagctttatgaacaccaacctg gaaaacaacaaaaaagtgctgattgaaggcgcgaacgcggcgatgctggatattgatttt ggcacctatccgtatgtgaccagcagctgcaccaccgtgggcggcgtgtttagcggcctg ggcattcatcataaaaaactgaacctggtggtgggcgtggtgaaaagctatctgacccgc gtgggctgcggcccgtttctgaccgaactgaacaacgatgtgggccagtatctgcgcgaa aaaggccatgaatatggcaccaccaccaaacgcccgcgccgctgcggctggctggatatt ccgatgctgctgtatgtgaaatgcattaacagcattgatatgattaacctgaccaaactg gatgtgctgagcggcctggaagaaattctgctgtgcgtgaactttaaaaacaaaaaaacc ggcgaactgctggaaaaaggctgctatccggtggaagaagaaattagcgaagaatatgaa ccggtgtatgaaaaatttagcggctggaaagaagatattagcacctgcaacgaatttgat gaactgccggaaaacgcgaaaaaatatattctggcgattgaaaaatatctgaaaaccccg attgtgtggattggcgtgggcccgaaccgcaaaaacatgattgtgaaaaaaaactttaac ctgaac
 * TMpred graph Image ** (http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.
 * *CDS ** ** Gene Sequence ** ** (paste as text only): **


 * *GC% Content for gene: 41% **

Do Not Need this info for Spring (but still copy these lines to your Target page for now) ***GC% Content for gene (codon optimized):** **Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers):** **(**** link to DNA Works output ** ** text file ** ** - **that should be saved in your Google Docs folder after you did the primer design protocol) -- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to. **Primer design results for 'tail' primers (this is just 2 sequences):**
 * *CDS Gene Sequence (codon optimized) - copy from ** ** output ** ** of ** ** Primer Design ** ** Protocol (paste as text only): **