TargetSp15+-+Glucose-1-Phosphatase+(Salmonella+Enterica)

Salmonella is among the most common food-borne diseases, causing approximately 380 deaths per year in the US. Those with particularly vulnerable immune systems like children and the elderly are most prone to this disease. Salmonella enterica is one of the two most common strains of salmonella, along with S. Typhimurium. "The presence of a hexose phosphatase with similar relative specificity in the periplasmic space of Salmonella typhimurium has been reported (25). The ability of E. coli strains lacking the hexose phosphate transport system to utilize glucose-i-phosphate (G-1-P) as the sole carbon source has also been considered as an indication that some periplasmic phosphatase could efficiently cleave G-1-P (13). Inactivating agp on the chromosome of a wild-type strain indicated that GlPase was indeed absolutely required for the growth of E. coli in a high-phosphate medium containing G-1-P as the sole carbon source" paper]
 * *Target (protein/gene name): **Glucose-1-phosphatase
 * *NCBI Gene # or RefSeq#: ** 1247561
 * *Protein ID (NP or XP #) or Wolbachia#: **NP_455612. 1
 * *Organism (including strain): **Salmonella Enterica (str. 08-0047)
 * Etiologic Risk Group (see link below): ** Category B Priority Pathogens
 * */ Disease Information (sort of like the Intro to your Mini __Research Write__ up): **
 * 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: **Used in glycolysis and glu con eogenesis pathways, essential for energy synthesis. Important in initial pathway for glycolysis, however, alternate routes available.

Is it a monomer or multim er as biological unit ** ? (make prediction at ** @http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html): Two amino acid chains and two ligands in ASU. Likely a multimer.

Inhibitors found: Phosphate (product inhibition) [3,4]; F- [4], Arsenate [4] paper] --BRENDA enzyme mechanism schematic
 * Complex of proteins?: **No
 * Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): **
 * *EC#: ** 3.1.3.10
 * Link to BRENDA EC# page: **

Enzyme assay with pNPP as substrate, with potassium phalate buffer (E. coli) paper]
 * Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): **
 * -- links to assay reagents (substrates) pages. **
 * --- List cost and quantity of substrate reagents, supplier, and catalog # **

"Glucose-1-phosphatase assays were performed in 200 ml of 0.1 M sodium acetate buffer pH 4.5 using 5 mM of glucose-1- phosphate as the substrate at 310 K for 10 min. The reaction was stopped by adding 133 ml of a mixture of ammonium molybdate, ammonium vanadate and nitric acid (Engelen et al., 1994). The concentration of inorganic phosphate released was determined by comparison with a K2HPO4 standard measured at 405 nm." paper]

-- PDB # or closest PDB entry if using homology model: 1NT4 -- For Homology Model option: Show pairwise alignment of your BLASTP search in NCBI against the PDB Query Coverage: 94% Max % Identities: 83% % Positives: 90% Chain used for homology: QTVPEGYQLQQVLMMSRANLRAPLANNGSVLEQSTPNKWPEWDVPGGQLTTKGGVLEVYMGHYMREWLAE
 * Structure (PDB or Homology model) **

QGMVKSGECPPPYTVYAYANSLQRTVATAQFFITGAFPGCDIPVHHQEKMGTMDPTFNPVITDDSAAFSE

QAVAAMEKELSKLQLTDSYQLLEKIVNYKDSPACKEKQQCSLVDGKNTFSAKYQQEPGVSGPLKVGNSLV

DAFTLQYYEGFPMDQVAWGEIKSDQQWKVLSKLKNGYQDSLFTSPEVARNVAKPLVSYIDKALVTDRTSA

PKITVLVGHDSNIASLLTALDFKPYQLHDQNERTPIGGKIVFQRWHDSKANRDLMKIEYVYQSAEQLRNA DALTLQAPAQRVTLELSGCPIDADGFCPMDKFDSVLNEAVK

Phosphate (product inhibition) [3,4]; F- [4], Arsenate [4]
 * Current Inhibitors: **

Structural gene AGP can be cloned to overexpress E. coli phosphatases, including Glucose-1-phosphatase. paper, [|paper]] "Glucose-1-phosphatase was purified from the osmotic shock fluid of a 1-liter culture of strain SBS1405(pEP1376) grown to early stationary phase in TYE medium plus ampicillin. Low-molecular-weight proteins were removed, and the enzyme was concentrated with an Amicon concentrator equipped with an UM20 membrane. The concentrated preparation was chromatographed on a fast protein liquid chromatography Mono-Q column (Pharmacia) in 5 mM Tris hydrochloride buffer (pH 7.8) with a linear gradient of NaCl (0 to 500 mM)." [[|paper]]
 * Expression Information (has it been expressed in bacterial cells): **
 * Purification Method : **
 * Image of protein (PyMol with features delineated and shown separately): **
 * *Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): **

MKKSLLAVAVAGAVLLSSAVQAQTTPEGYQLQQVLMMSRHNLRAPLANNGNVLAQSTPNAWPAWDVPGGQ

LTTKGGVLEVYMGHYTREWLVAQGLIPSGECPAPDTVYAYANSLQRTVATAQFFITSAFPGCDIPVHHQE

KMGTMDPTFNPVITDDSAAFRQQAVQAMEKARSQLHLDESYKLLEQITHYQDSPSCKEKHQCSLIDAKDT

FSANYQQEPGVQGPLKVGNSLVDAFTLQYYEGFPMDQVAWGGIHTDRQWKVLSKLKNGYQDSLFTSPTVA

RNVAAPLVKYIDKVLVAERVSAPKVTVLVGHDSNIASLLTALDFKPYQLHDQYERTPIGGQLVFQRWHDG

NANRDLMKIEYVYQSARQLRNAEALTLKSPAQRVTLELKGCPVDANGFCPLDKFDNVMNTAAK
 * *length of your protein in Amino Acids: **413
 * Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website: **45629.85 kD
 * Molar Extinction coefficient of your protein at 280 nm wavelength: ** 56840
 * 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): **