Target+-+S-adenosylhomocysteine+hydrolase+(L.+major)


 * Target (protein/gene name):** S-adenosylhomocysteine hydrolase
 * NCBI Gene # or RefSeq#:** 5655685 (full gene) or 223365960 (Chain A)
 * Protein ID (NP or XP #) or Wolbachia#:** XP_001686974.1; Gene symbol: LmjF36.3910
 * Organism (including strain):** Leishmania major strain Friedlin, chain A
 * Etiologic Risk Group (see link below):** 2 ([])

Initiated by the bite of a sandfly, leishmaniasis is a tropical disease characterized by skin sores and damage of internal organs (1). Although noticeable symptoms may not appear at first, after an incubation of several weeks or months cutaneous leishmaniasis can cause bumps on the skin (cutaneous leishmaniasis). The infection can then develop rapidly, turning into volcanic sores that are raised on the outside with a crater in the center (1). Typically, these sores do not cause severe pain. The more serious side of the infection (visceral leishmaniasis) includes the enlarging of certain organs, such as the liver and spleen, and can effect bone marrow. Low red blood cell count due to the disease can also cause anemia, which takes several months or years to develop (2). Currently, leishmaniasis is categorized as a tropical disease that is lacking in effective control measures (3). The disease is a vector-borne, caused by the parasitic protozoan //Leishmania major.// The high reproductive potential of //L. major// allows it to recover quickly after being treated with pharmaceutical drugs, which hinders global eradication programs (3). The protozoa are transferred from the gut of the sandfly to the human host once the human is bitten. The chosen drug target, S-adenosylhomocysteine hydrolase, is important in the cysteine-methionine and selenoamino acid metabolic pathways in the //L. major// parasite (4).
 * Background/ Disease Information (sort of like the Intro to your Mini Research Write up):**

References: 1. [] 2. [] 3. [] 4. []


 * Link to TDR Targets page (if present):** []
 * Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.):** []
 * Essentiality of this protein:** Yes
 * 1. Gene/Ortholog:** Tb11.01.1350 (OG4_10673); **Phenotype:** significant gain of fitness in bloodstream forms (3 days); **Source study:** alsford
 * 2. Gene/Ortholog:** Tb11.01.1350 (OG4_10673); **Phenotype:** significant gain of fitness in bloodstream forms (6 days); **Source study:** alsford
 * 3. Gene/Ortholog:** Tb11.01.1350 (OG4_10673); **Phenotype:** no significant loss or gain of fitness in procyclic forms; **Source study:** alsford
 * 4. Gene/Ortholog:** Tb11.01.1350 (OG4_10673); **Phenotype:** no significant loss or gain of fitness in differentiation of procyclic to bloodstream forms; **Source study:** alsford
 * Complex of proteins?:** 4 chains; Protein-ligand complex; []
 * Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):** 0.8 (according to TDR Targets)


 * *EC#: 3.3.1.1 **
 * Link to BRENDA EC# page:** []
 * Figure1.** Screenshot of BRENDA enzyme mechanism schematic

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 * Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):**
 * Assay Reagent/Substrate:** S-adenosylhomocysteine (S-AdoHcy) with ThioGlo1
 * Assay Method:** Fluorescent
 * Supplier:** Sigma-Aldrich
 * Cost:** $63.70 for 10mg of substrate.............$594 for 1 mg ThioGlo
 * CAS:** Substrate 979-92-0/ThioGlo 168639-87-0
 * Article:** []
 * Reagent links:**

PDB #: **3G1U**
 * Structure Available (PDB or Homology model)**

[|CHEMBL1095280], [|CHEMBL1098945] , [|CHEMBL8771] , [|CHEMBL155085] , [|CHEMBL1097885] BindingDB.org Article ID: 50031613 [] "Compounds that specifically inhibit S-adenosylhomocysteine hydrolase (SAHH; EC 3.3.1.1) interfere with the proliferation of Plasmodium malarial parasites, but efforts to identify the enzyme directly in parasite extracts have been unsuccessful. Here we report genetic and biochemical evidence for the presence of a gene encoding P. falciparum SAHH. The gene is transcribed as a 2.8-kilobase mRNA in erythrocytic stage parasites. Analysis of the open reading frame predicts a 53.9-kDa protein having conserved regions thought to be involved in NAD binding. The cDNA sequence has been incorporated into an Escherichia coli expression construct to confirm the function of the sahh product ." Article: //Plasmodium falciparum S-adenosylhomocysteine hydrolase. cDNA identification, predicted protein sequence, and expression in Escherichia coli.// [] "S-Adenosyl-L-homocysteine hydrolase has been purified to apparent homogeneity from rat liver by means of affinity chromatography on 8-(3-aminopropylamino)adenosine linked to Sepharose." [] "SAH hydrolase was purified to apparent homogeneity from bovine kidney by standard chromatographic methods. SAH hydrolase was converted in its reduced form, which had the advantage that the SAH hydrolase is enzymatically inactive." [|http://go.galegroup.com/ps/retrieve.do?sgHitCountType=None&sort=RELEVANCE&inPS=true&prodId=AONE&userGroupName=txshracd2598&tabID=T002&searchId=R1&resultListType=RESULT_LIST&contentSegment=&searchType=AdvancedSearchForm&currentPosition=1&contentSet=GALE%7CA209407238&&docId=GALE|A209407238&docType=GALE&role=]
 * Current Inhibitors:**
 * Expression Information (has it been expressed in bacterial cells):** Yes
 * Purification Method :**


 * Image of protein (PyMol with features delineated and shown separately):**
 * Figure 2.** 3G1U protein shown as cartoon colored by chain.
 * Figure 3.** 3G1U protein chains shown as sticks colored with carbons green, nitrogen blue, and oxygen red.

>3G1U:A|PDBID|CHAIN|SEQUENCE MADYKVKDISLAEWGRKAIELAENEMPGLMELRREYGPSQPLKGAKIAGCLHMTVQTAVLIETLKALGAELRWSSCNIFS TQDNAAAAIAKTGVPVFAWKGETDEEYEWCIAQTVKGFSGDGLPNMILDDGGDLTNLVIDRYPELVPKIFGISEETTTGVKNLYKRLSKGNLPISAINVNDSVTKSKFD NLYGCRESLVDGIKRATDVMIAGKTCCVCGYGDVGKGCAAALRAFGARVVVTEVDPINALQASMEGYQVALVEDVMADAHIFVTTTGNDDIITSDHFPHMRDDAIVCN IGHFDTEIQVGWLEANAKEHVEIKPQVDRYTMENGRHIILLAKGRLVNLGCASGHPSFVMSNSFTNQVLAQIELWSNRDNGKYPRGDKAGVFFLPKALDEKVAALHL AHVGAKLTKLTPKQAEYINCPVNGPFKPDHYRY (*sequence for Chain A only) code Ext. coefficient   52995 Abs 0.1% (=1 g/l)  1.108, assuming all pairs of Cys residues form cystines
 * Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):**
 * length of your protein in Amino Acids:** 437 amino acids
 * Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website:** 47828.7
 * Molar Extinction coefficient of your protein at 280 nm wavelength:**

Ext. coefficient   52370 Abs 0.1% (=1 g/l)  1.095, assuming all Cys residues are reduced code atggcggact acaaggtaaa agacatcagc ctcgcggagt ggggccgcaa ggcgatcgag ctcgcggaaa acgagatgcc cggtctgatg gagctgcgcc gcgagtacgg accctcccag ccgctgaagg gcgccaagat tgccggctgc ctgcacatga ccgtacagac tgccgtgctg atcgagaccc tcaaggccct cggtgcggag ctgcgctggt cgtcgtgcaa catcttttcc actcaggata acgccgctgc ggccattgcc aagactggcg taccggtgtt tgcgtggaag ggtgagacag acgaggagta tgagtggtgc atcgcccaga cagtgaaggg cttcagcggt gacggtctgc cgaacatgat cctcgacgac ggtggcgacc tcacgaatct ggtgatcgac cgctaccccg agctcgtgcc caagatcttc ggtatctccg aggagaccac gacgggtgtg aagaacctgt acaagcgcct gagcaagggc aacctcccca tctccgccat caacgttaac gacagcgtga cgaagagcaa gttcgacaac ctttacggct gccgagagtc cttggtggac ggcatcaagc gcgccacgga tgtcatgatt gccggcaaga cgtgctgcgt gtgcggatac ggtgatgttg gtaagggctg cgccgccgcc ctgcgcgcct tcggcgctcg cgttgtggta acggaggtgg accctatcaa tgcccttcag gcctccatgg agggctacca ggtcgctctg gtcgaagacg tcatggccga tgcgcacatc ttcgtgacga ccaccggcaa cgatgacatc atcacctctg accacttccc tcacatgcgc gacgacgcca ttgtgtgcaa catcggccac ttcgacacgg agatccaggt gggatggctt gaggcaaacg ccaaagagca cgtggaaatc aagcctcagg tggaccgcta taccatggag aacggccgtc acatcatcct gctggctaag ggccgcctgg tcaacctcgg ctgcgccagc ggtcacccgt ccttcgtgat gtccaactcc ttcacgaacc aggtgctggc tcagatcgag ctctggagca accgcgacaa cggcaagtac ccgcgcggtg acaaggccgg cgtgttcttc ctgcccaagg cgctcgatga gaaggtcgcc gccctccacc tcgcccacgt cggcgccaag ctgacaaaac tgaccccgaa gcaggccgag tacatcaact gcccggtcaa cggcccgttc aagccggacc actaccgcta ctaa
 * TMpred graph Image ** (@http://www.ch.embnet.org/software/TMPRED_form.html).
 * CDS Gene Sequence (paste as text only):**

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 * GC% Content for gene:**


 * CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):**
 * GC% Content for gene (codon optimized):**

Do Not Need this info for Spring (but still copy these lines to your Target page for now) -- 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 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)


 * Primer design results for 'tail' primers (this is just 2 sequences):**