Target+-+Serine+Threonine+Phosphatase+STP1,+(Streptococcus+Agalactiae)


 * Target Team:** Jairo V, Saniya H, Stacy T, Daren
 * Target (protein/gene name):** Serine Threonine Phosphatase, STP1 SAK_0388
 * NCBI Gene #:** 3686217
 * Protein ID (NP or XP #):** YP_329040
 * Organism (including strain):** Streptococcus Agalactiae, Strain A909
 * Etiologic Risk Group**: Risk Group 2
 * Disease Information:** Streptococcus Agalactiae is a bacterial infection that is mostly found in the intestinal tracts of humans, but it can also reside in the genital tracts of women. If a woman is infected with the bacteria, it can be transferred to the newborn baby through the amniotic fluid during childbirth. The newborn baby would then either develop the early or late onset disease which have the symptoms of respiratory distress, fever, lethargy, irritability, apnea and hypotension, but the late onset disease also causes meningitis and bacterimia. Only 2% of all the babies born are infected with this bacteria, and out all of those, more than 60% of the newborn babies become infected within the first 24 hours, and only 8% become infected after 2 days.The mortality rate, even with current treatments, exceeds 20%, and it becomes twice as deadly on older infants. It also affects people who are older than 65, with a 15% mortality rate, and adults who suffer alcoholism, diabetes, HIV, etc. Streptococcus Agalactiea is susceptible to combinations of ampicillin and penicillin,vancomycin, ciprofloxacin, clindamycin, erythromycin, cotrimoxazole, ceftriaxone, and it is resistant to macrolides and clindamycin.
 * Image 1. Individual colony of Streptococcus Agalactiae.**

http://www.uniprot.org/uniprot/Q3K363
 * Potential Physiological Targets:** GST-Stk1 & PpaC (via: http://www.phosphatase.biochem.vt.edu/p3d2.cfm?extf=accession&extref=P3D0411151)
 * Link to Gene Database page:** http://www.ncbi.nlm.nih.gov/gene/3686217
 * Is it a monomer or multimer as biological unit? (prediction):** monomer


 * EC#:** 3.1.3.16
 * Link to BRENDA EC# page:** http://www.brenda-enzymes.org/enzyme.php?ecno=3.1.3.16
 * FIgure 1. The Brenda enzyme mechanism schematic for the reaction of STP1, a phosphoprotein, and water.**


 * Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):**
 * link to Sigma (or other company) page for assay (see Sigma links below)**
 * 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 #**

-- For Homology Model option:
 * Structure (PDB or Homology model)**
 * PDB #:** 2PK0
 * Ligands:** Chloride Ions, Glycerol and Magnesium Ions.
 * Show pairwise alignment of your BLASTP search in NCBI against the PDB**
 * Query Coverage: 100%**
 * Max % Identities: 99%**
 * % Positives: 244/245 (99%)**
 * Chain used for homology: A, B, C, D**




 * Current Inhibitors:** Okadaic Acid, Microcystin LR Cantharidin, Inhibitor-2, EDTA & FK-506.
 * Expression Information (has it been expressed in bacterial cells):**
 * Purification Method:**
 * Image of protein:**
 * Image 2. The PyMol image of the STP1 protein, with the main chain carbons colored green and shown as ribbons, the magnesium ions shown as red spheres, the chlorine ions shown as yellow spheres, the glycerol shown as magenta and the active site cyan.**

1 MEISLLTDIGQRRSNNQDFINQFENKAGVPLIILADGMGGHRAGNIASEMTVTDLGSDWA
 * Amino Acid Sequence:**

61 ETDFSELSEIRDWMLVSIETENRKIYELGQSDDYKGMGTTIEAVAIVGDNIIFAHVGDSR

121 IGIVRQGEYHLLTSDHSLVNELVKAGQLTEEEAASHPQKNIITQSIGQANPVEPDLGVHL

181 LEEGDYLVVNSDGLTNMLSNADIATVLTQEKPLDDKNQDLITLANHRGGLDNITVALVYV

241 ESEAVX


 * Length of your protein in Amino Acids:** 246 amino acids


 * Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website:** 26840 kDA


 * Molar Extinction coefficient of your protein at 280 nm wavelength:** 18450 M -1 cm -1


 * TMpred graph Image (@http://www.ch.embnet.org/software/TMPRED_form.html):**

code ATGGAAATATCTCTTTTAACAGATATTGGGCAACGTCGTTCCAATAATCAAGATTTTATTAATCAATTTG AAAATAAAGCCGGCGTCCCACTAATCATCTTAGCAGATGGTATGGGAGGACATCGTGCAGGAAATATTGC TAGCGAGATGACTGTAACTGATCTAGGAAGTGATTGGGCGGAGACAGATTTTTCTGAGTTATCAGAAATT CGTGATTGGATGCTTGTCTCTATTGAAACAGAGAATCGCAAAATATATGAATTAGGTCAGTCAGATGATT ACAAAGGTATGGGAACTACCATTGAAGCAGTTGCCATTGTAGGAGATAATATTATTTTCGCACATGTAGG AGATTCACGTATCGGTATCGTTCGTCAAGGAGAGTATCATTTATTGACGAGTGATCATTCTCTTGTTAAT GAATTGGTAAAAGCTGGCCAATTAACAGAAGAAGAAGCTGCTAGTCATCCTCAAAAAAATATTATTACGC AATCAATTGGTCAAGCCAATCCAGTCGAACCTGATTTAGGAGTTCATCTACTTGAAGAAGGTGACTACCT TGTAGTTAATAGTGATGGACTAACTAATATGTTGAGTAATGCCGATATTGCTACAGTACTAACACAAGAA AAAACCTTAGATGATAAGAATCAAGATTTAATCACTCTAGCAAACCATAGAGGAGGTCTTGATAATATCA CTGTGGCACTAGTTTACGTTGAAAGTGAGGCGGTTTAA code
 * CDS Gene Sequence (paste as text only):**
 * GC% Content for gene:** 37.1%

1 ATGGAGATCAGCCTGCTCACTGACATTGGCCAGCGTCGTTCTAACAATCAGGACTTCATC 61 AATCAGTTCGAAAACAAAGCGGGTGTTCCGCTGATCATCCTGGCGGACGGCATGGGCGGT 121 CACCGTGCGGGCAACATCGCTTCCGAAATGACCGTTACCGACCTGGGTTCCGACTGGGCG 181 GAGACCGACTTCTCTGAACTGTCTGAAATTCGTGACTGGATGCTGGTATCTATCGAAACC 241 GAAAACCGTAAAATCTACGAACTCGGTCAATCTGACGACTACAAAGGTATGGGTACCACC 301 ATCGAAGCCGTAGCGATCGTTGGTGACAATATCATCTTCGCGCACGTCGGTGACTCTCGT 361 ATCGGTATCGTTCGTCAAGGTGAATATCACCTGCTGACGAGCGACCACTCTCTGGTTAAC 421 GAACTGGTCAAAGCAGGCCAGCTGACTGAAGAGGAAGCGGCGTCTCACCCGCAGAAAAAC 481 ATCATCACCCAGTCTATCGGTCAGGCGAATCCGGTTGAACCGGACCTCGGTGTACATCTG 541 CTGGAAGAAGGTGACTACCTGGTTGTTAACTCTGACGGTCTGACCAACATGCTGTCTAAC 601 GCGGACATCGCGACCGTTCTGACCCAGGAAAAACCGCTGGACGACAAAAACCAGGACCTG 661 ATCACCCTGGCGAACCATCGTGGTGGTCTGGACAACATCACCGTTGCGCTGGTTTACGTT 721 GAATCTGAAGCGGTATAA
 * CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):**


 * GC% Content for gene (codon optimized):** 52.8%

(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 pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers):**

https://docs.google.com/document/d/1ib30OT6lGE97Oa7ta5o1mlVMiuNLtMUCS9A8EqAKTPI/edit


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