Target-MtnX+phosphatase+(Bacillus+Anthracis)

1088838 Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Bacillus; Bacillus cereus group. ***Background/Disease Information** (sort of like the Intro to your Mini Research Write up):
 * *Target (protein/gene name): ** MtnX Phospatase
 * *NCBI Gene # or RefSeq#: ** 30264114
 * *Protein ID (NP or XP #) or Wolbachia#: **NP_846491.1
 * *Organism (including strain): ** Bacillus anthracis str. Ames
 * Etiologic Risk Group (see link below): **Appendix B-II-A. Risk Group 2 (RG2) - Bacterial Agents

Bacillus anthracis is the cause or origin of the disease commonly known as anthrax. B.anthracis is a rod shaped bacterium, that forms endospores and is gram-positive. It can be grown in aerobic or anaerobic conditions and is one of the few bacterium's that is known to synthesize the protein capsule, poly-Y-D-gamma-glutamic acid.This acid capsule of Bacillus anthracis is the lethal toxin agent found in anthrax [1]. The PGA capsule is able to avoid immune cell regulation and can proliferate within the host, making it an important factor in the pathogenesis of anthrax infection. MtnX plays an important role in the function of this organism because it catalyzes the dephosphorylation of 2-hydroxy-3-keto-5-methylthinopentenyl-1-phosphate into mtnD [2]. This MtnX phosphatase contributes to energy supply and regulation within Bacillus anthracis that is necessary for the bacterium to function.

Essentiality of this protein: Essential http://onlinelibrary.wiley.com/doi/10.1002/prot.21602/full Complex of proteins?: No Druggable Target: Yes

Information for the mechanism schematic could not be found for organism Bacillus anthracis on BRENDA. However, MtnX functions similarly in organism Bacillus subtilis.
 * *EC#: ** 3.1.3.87
 * Link to BRENDA EC# page: ** 3.1.3.87

p-Nitrophenyl Phosphate Liquid Substrate System pNPP does not work in Klebsiella pneumoniae version Crystal structure of MtnX phosphatase from Bacillus subtilis at 2.0 Å resolution provides a structural basis for bipartite phosphomonoester hydrolysis of 2-hydroxy-3-keto-5-methylthiopentenyl-1-phosphate N7653-100ML, $ 79.90 (Sigma-Aldrich, USA) MtnX Phosphatase with Organism Bacillus anthracis is not available on the PDB webpage. However, MtnX Phosphatase with organism Bacillus subtilis is provided and the information for that is included below. It is recommend that a Homology model is created for this phosphatase. -- __PDB # or closest PDB entry if using homology model:__ 2FEA -- __For Homology Model__: Will be required.
 * Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): ** spectrophotometic
 * -- link to Sigma (or other company) page for assay or assay reagents (substrates): **
 * -- link (or citation) to paper that contains assay information: **
 * -- List cost and quantity of substrate reagents and supplier: **
 * Structure Available (PDB or Homology model) **

__Query Coverage:__ 97% __Max % Identities:__ 52% __% Positives:__ 167/214(78%) __Chain used for homology FASTA__: **Protein Database (2FEA) for protein sequence MtnX in Bacillus subtilis that will be used for homology model:** >2FEA:A|PDBID|CHAIN|SEQUENCE GMTTRKPFIICDFDGTITMNDNIINIMKTFAPPEWMALKDGVLSKTLSIKEGVGRMFGLLPSSLKEEITSFVLEDAKIRE GFREFVAFINEHEIPFYVISGGMDFFVYPLLEGIVEKDRIYCNHASFDNDYIHIDWPHSCKGTCSNQCGCCKPSVIHELS EPNQYIIMIGDSVTDVEAAKLSDLCFARDYLLNECREQNLNHLPYQDFYEIRKEIENVKEVQEWLQNKNAGESSLK >gi|30264114|ref|NP_846491.1| 2-hydroxy-3-keto-5-methylthiopentenyl-1-phosphate phosphatase [Bacillus anthracis str. Ames] MSIQVFCDFDGTITNNDNIMSIMEKFAPPEAEEVKNRILSQELSIQEGVSQLFQLIPTNLHDEIIQFLIE TAEIRNGFHEFIQFVNENNISFYVISGGMDFFVYPLLQGLIPKEQIYCNETDFSNEYITVNWPHPCDRLC QNHCGLCKSSLIRKLSDTNDFHIVIGDSITDLQAAKQADKVFARDFLITKCEENHISYTPFETFHDVKTE LKHLLEVKL
 * NCBI Reference Sequence (NP_846491.1) protein sequence MtnX in Bacillus anthracis:**

MSIQVFCDFDGTITNNDNIMSIMEKFAPPEAEEVKNRILSQELSIQEGVSQLFQLIPTNLHDEIIQFLIE TAEIRNGFHEFIQFVNENNISFYVISGGMDFFVYPLLQGLIPKEQIYCNETDFSNEYITVNWPHPCDRLC QNHCGLCKSSLIRKLSDTNDFHIVIGDSITDLQAAKQADKVFARDFLITKCEENHISYTPFETFHDVKTE LKHLLEVKL Ext. coefficient 13325 Abs 0.1% (=1 g/l) 0.527, assuming all pairs of Cys residues form cystines (with Cys) Ext. coefficient 12950 Abs 0.1% (=1 g/l) 0.512, assuming all Cys residues are reduced (without Cys)
 * Current Inhibitors:** N/A
 * Expression Information (has it been expressed in bacterial cells):** Escherichia coli
 * Purification Method:** N/A
 * Image of protein (PyMol with features delineated and shown separately): **
 * *Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): **
 * *length of your protein in Amino Acids: ** 219aa
 * Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam]website: **25289.7 kDa
 * Molar Extinction coefficient of your protein at 280 nm wavelength: **
 * TMpred graph Image ** (@http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.

ATGAAACGTATTAAAATTTCAACAGAGTATATTACACTAGGGCAATTTTTAAAGTTAGCCGATGTAATTG ATACAGGTGGCGCTGTAAAATGGTTTTTACAAGAATATGAAGTGTACGTGAATCAAGAACTTGAAAATAG AAGAGGGCGCAAGCTATATGCGAACGATATTATTGAAATTCCAGGAAGCGGAAGTTTCCAAGTTCAGTCA TAA
 * *CDS Gene Sequence (paste as text only): **
 * *GC% Content for gene: ** 34.74%
 * *CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only): N/A (not needed currently) **
 * *GC% Content for gene (codon optimized): N/A (not needed Currently) **

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


 * References: **

[1] Jang J., The poly-y-D-glutamic acid capsule of Bacillus anthracis enhances lethal toxin activity. //Epub// **2011**, 79 (9), 3846-54.

[2] Xu Q., Crystal structure of MtnX phosphatase from Bacillus subtillis at 2.0 angstroms resolution provides a structural basis for bipartite phosphomonoester hydrolysis of 2-hydroxy-3-keto-5-methylthiopentenyl-1-phosphate. //PubMed// **2007**, 69 (2), 433-9.