TargetSp+15+-+E3+Ubiquitin-protein+ligase+(Salmonella+Typhimurium)


 * *Target (protein/gene name): ** E3 ubiquitin-protein ligase/sspH1
 * *NCBI Gene # or RefSeq#: ** 888858 [NCBI] RefSeq#: YP_005237063.1.NC_016856.1.
 * *Protein ID (NP or XP #) or Wolbachia#: **NC_020282.1
 * *Organism (including strain): ** Salmonella typhimurium (strain 14028s / SGSC 2262)


 * Etiologic Risk Group (see link below): ** Risk group lv 2 (moderate individual risk, low community risk)

Many types of Salmonella bacteria cause salmonellosis in animals and people. While the occurrence of different types of Salmonella varies from country to country, S. typhimurium and S. enteritidis are the two most commonly found in the United States.
 * */ Disease Information (sort of like the Intro to your Mini __Research Write__ up): **

Some strains of Salmonella have become resistant to several antibiotics normally used to treat people with salmonella disease, posing a serious public health threat.

Every year, Salmonella is estimated to cause one million illnesses in the United States, with 19,000 hospitalizations and 380 deaths. Most persons infected with Salmonella develop diarrhea, fever, and abdominal cramps 12 to 72 hours after infection.

The illness usually lasts 4 to 7 days, and most persons recover without treatment. However, in some persons, the diarrhea may be so severe that the patient needs to be hospitalized.


 * Link to TDR Targets page (if present): ** Not available

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 * Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.): **


 * Essentiality of this protein: **The protein interferes with the host's ubiquitination pathway and targets host proteins for proteasomal degradation

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?: **
 * Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): **

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 * *EC#: ** 6.3.2.19
 * Link to BRENDA EC# page: **


 * -- ** Show screenshot of BRENDA enzyme mechanism schematic



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 * Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): **
 * -- link to Sigma (or other company ) page for assay (see Sigma links below): **
 * -- -or link (or citation) to paper that contains assay information: **
 * -- links to assay reagents (substrates) pages. **
 * --- List cost and quantity of substrate reagents, supplier, and catalog # **

-- PDB # or closest PDB entry if using homology model: -- For Homology Model option: Show pairwise alignment of your BLASTP search in NCBI against the PDB Query Coverage: Max % Identities: % Positives Chain used for homology:
 * Structure (PDB or Homology model) **

Protein constructs were expressed in BL21 (DE3) CodonPlus and induced with 0.3 mm isopropyl-1-thio-β-d-galactopyranoside at 16 °C for overnight. Expressed proteins were purified using nickel-nitrilotriacetic acid affinity chromatography and stored in a buffer containing 20 mmHEPES (pH 7.5), 150 mm NaCl, 5% glycerol, and 1 mm DTT. Purification tags on UBE1, UBE2D3, and ubiquitin were removed by tobacco etch virus protease.
 * Current Inhibitors: ** paH enzymes
 * Expression Information (has it been expressed in bacterial cells): ** Yersinia pestis
 * Purification Method : **
 * Image of protein (PyMol with features delineated and shown separately): **


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MFNIRNTQPS VSMQAIAGAA APEASPEEIV WEKIQVFFPQ ENYEEAQQCL
 * *Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): **

AELCHPARGM LPDHISSQFA RLKALTFPAW EENIQCNRDG INQFCILDAG

SKEILSITLD DAGNYTVNCQ GYSEAHDFIM DTEPGEECTE FAEGASGTSL

RPATTVSQKA AEYDAVWSKW ERDAPAGESP GRAAVVQEMR DCLNNGNPVL

NVGASGLTTL PDRLPPHITT LVIPDNNLTS LPELPEGLRE LEVSGNLQLT

SLPSLPQGLQ KLWAYNNWLA SLPTLPPGLG DLAVSNNQLT SLPEMPPALR

ELRVSGNNLT SLPALPSGLQ KLWAYNNRLT SLPEMSPGLQ ELDVSHNQLT

RLPQSLTGLS SAARVYLDGN PLSVRTLQAL RDIIGHSGIR IHFDMAGPSV

PREARALHLA VADWLTSARE GEAAQADRWQ AFGLEDNAAA FSLVLDRLRE

TENFKKDAGF KAQISSWLTQ LAEDAALRAK TFAMATEATS TCEDRVTHAL

HQMNNVQLVH NAEKGEYDNN LQGLVSTGRE MFRLATLEQI AREKAGTLAL

VDDVEVYLAF QNKLKESLEL TSVTSEMRFF DVSGVTVSDL QAAELQVKTA

ENSGFSKWIL QWGPLHSVLE RKVPERFNAL REKQISDYED TYRKLYDEVL

KSSGLVDDTD AERTIGVSAM DSAKKEFLDG LRALVDEVLG SYLTARWRLN


 * *length of your protein in Amino Acids: ** 700
 * Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website: ** 77,119
 * Molar Extinction coefficient of your protein at 280 nm wavelength: ** 91370
 * 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: **
 * *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): **