Target+-+1-DEOXY-D-XYLULOSE+5-PHOSPHATE+REDUCTOISOMERASE+DXR+(Mycobacterium+Tuberculosis)


 * *Target (protein/gene name): 1-deoxy-D-xylulose-5-phosphate reductoisomerase **


 * *NCBI Gene # or RefSeq#: **887800

***Locus Tag:** RV2870C


 * *Protein ID (NP or XP #) or Wolbachia#: **NP_217386.2


 * *Organism (including strain): Mycobacteria Tuberculosis **


 * Etiologic Risk Group (see link below): Risk Group 2 **

Mycobacterium Tuberculosis organisms are obligate aerobes growing most successfully in tissues abundant with oxygen, such as the lungs. They are considered as "facultative intracellular pathogens" and usually affect mononuclear phagocytes such as macrophages. Physically, they are hydrophobic and have a high lipid content in the cell wall. Because the cells are hydrophobic and tend to clump together, they are impervious to classic staining techniques (Gram's stain). The disease caused by the organism, Tuberculosis, generally includes an onset of symptoms containing fever, weight-loss, constant cough, and lethargy. Today TB is prevented in most developed countries via vaccination at birth. If an individual has not garnered immunity and is infected at a later point, drugs such as INH, rifampin, pyrazinamide, or ethmbutol may be used.
 * *Background/Disease Information (sort of like the Intro to your Mini Research Write up): **

"1-Deoxy-d-xylulose 5-phosphate reductoisomerase catalyzes the NADPH-dependent rearrangement and reduction of 1-deoxy-d-xylulose 5-phosphate to form 2-C-methyld-erythritol 4-phosphate, as the second step of the deoxyxylulose 5-phosphate/methylerythritol 4-phosphate pathway found in many bacteria and plants. The end product, isopentenyl diphosphate, is the precursor of various isoprenoids vital to all living organisms." ([] )
 * Essentiality of this protein: Essential. **


 * Complex of proteins?: No. **


 * Druggable Target: Yes. Drugability Index Score = .5 (TDR) **


 * *EC#: 1.1.1.267 **

1. [] 2. []
 * Link to BRENDA EC# page: **


 * Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): Spectrophotometric Assay (NADPH Absorption) **
 * -- 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: $379/kit. 400 assays in 96-well plates. **

Query Coverage: N/A Max % Identities: N/A
 * Structure Available (PDB or Homology model): Yes, PDB. PDB ID: 2Y1E **

% Positives: N/A Chain used for homology: N/A

List: []
 * Current Inhibitors: 33 Current Inhibitors **


 * Expression Information (has it been expressed in bacterial cells): Yes. **

The sequence positions in brackets denominate the core region.
 * Possible Transmembrane Helices**

 Only scores above 500 are considered significant. Inside to outside helices : 6 found from to score center 13 ( 15) 32 ( 32) 409 23 99 ( 99) 116 ( 116) 491 107  246 ( 246) 264 ( 264) 74 256  311 ( 311) 329 ( 329) 215 319  337 ( 337) 357 ( 355) 963 345  385 ( 385) 404 ( 402) 540 394

Outside to inside helices : 5 found from to score center 15 ( 15) 32 ( 32) 243 23 99 ( 99) 117 ( 117) 616 109  311 ( 311) 329 ( 329) 345 319  336 ( 336) 352 ( 352) 1020 344  382 ( 385) 403 ( 403) 230 395


 * Table of Correspondences**

Helices shown in brackets are considered insignificant.

 A "+"-symbol indicates a preference of this orientation.

 A "++"-symbol indicates a strong preference of this orientation.

inside->outside | outside->inside ( 13- 32 (20) 409 + ) |( 15- 32 (18) 243 ) ( 99- 116 (18) 491 ) | 99- 117 (19) 616 +  ( 246- 264 (19) 74 ++ ) |  ( 311- 329 (19) 215 ) |( 311- 329 (19) 345 + )  337- 357 (21) 963 | 336- 352 (17) 1020  385- 404 (20) 540 ++ |( 382- 403 (22) 230 )


 * Purification Method: Column chromatography. ( ** [] )


 * Image of protein (PyMol with features delineated and shown separately): **


 * *Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): **

1 mtnstdgrad grlrvvvlgs tgsigtqalq viadnpdrfe vvglaaggah ldtllrqraq 61 tgvtniavad ehaaqrvgdi pyhgsdaatr lveqteadvv lnalvgalgl rptlaalktg 121 arlalankes lvaggslvlr aarpgqivpv dsehsalaqc lrggtpdeva klvltasggp 181 frgwsaadle hvtpeqagah ptwsmgpmnt lnsaslvnkg leviethllf gipydridvv 241 vhpqsiihsm vtfidgstia qasppdmklp islalgwprr vsgaaaacdf htasswefep 301 ldtdvfpave larqagvagg cmtavynaan eeaaaaflag rigfpaivgi iadvlhaadq 361 wavepatvdd vldaqrware raqravsgma svaiastakp gaagrhastl ers


 * *length of your protein in Amino Acids: 413 Amino Acids **
 * Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website: 42853.4 g **
 * Molar Extinction coefficient of your protein at 280 nm wavelength: 37470 **
 * 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): **

ATGACTAACAGTACAGACGGTAGAGCTGATGGACGGTTGAGGGTAGTGGTTCTGGGGTCTACCGGGAGCATCGGT ACACAGGCCTTGCAAGTCATCGCAGACAATCCTGACCGATTCGAGGTGGTAGGCTTGGCAGCGGGTGGAGCGCAC CTAGATACACTCCTGCGGCAAAGGGCTCAGACTGGAGTTACGAATATCGCTGTCGCGGACGAACACGCCGCACAG AGGGTAGGGGACATCCCGTATCACGGCAGCGACGCTGCCACACGTCTGGTGGAGCAGACGGAAGCCGACGTCGT CCTCAACGCTTTAGTTGGTGCTCTAGGGCTCCGTCCAACGTTAGCTGCGCTAAAAACCGGCGCTCGTCTTGCATTAG CTAACAAGGAGTCCCTGGTAGCAGGTGGAAGCTTAGTACTCAGGGCCGCAAGACCTGGACAGATTGTACCGGTCG <span style="font-family: Arial,sans-serif; font-size: 10pt;">ATTCTGAGCATAGTGCACTCGCACAATGCCTACGCGGTGGAACCCCGGATGAAGTAGCGAAACTGGTATTAACTGC <span style="font-family: Arial,sans-serif; font-size: 10pt;">CAGTGGCGGGCCATTCAGAGGATGGTCCGCCGCTGACTTAGAACATGTGACACCAGAACAGGCCGGAGCTCATCC <span style="font-family: Arial,sans-serif; font-size: 10pt;">AACCTGGTCCATGGGCCCCATGAACACCCTGAATTCTGCAAGTTTGGTTAACAAAGGTTTAGAGGTGATAGAAACCC <span style="font-family: Arial,sans-serif; font-size: 10pt;">ACCTCTTATTCGGGATCCCTTACGACCGAATTGATGTAGTGGTACACCCTCAGTCCATAATACATTCGATGGTCACGT <span style="font-family: Arial,sans-serif; font-size: 10pt;">TCATCGATGGAAGTACTATCGCGCAGGCTTCACCACCGGATATGAAACTCCCGATATCTCTTGCGCTAGGATGGCCA <span style="font-family: Arial,sans-serif; font-size: 10pt;">CGTCGGGTGTCGGGCGCCGCGGCGGCTTGCGACTTCCACACAGCATCTTCCTGGGAATTTGAGCCGCTTGACACTG <span style="font-family: Arial,sans-serif; font-size: 10pt;">ACGTATCCCAGCCGTAGAGCTGGCACGTCAAGCGGGTGTTGCTGGGGGGTGCATGACCGCCGTATACAACGCTGC <span style="font-family: Arial,sans-serif; font-size: 10pt;">TAACGAAGAGGCTGCTGCCGCATTTTTAGCTGGGAGAATAGGCTTCCCCGCCATAGTCGGAATAATTGCTGATGTGC <span style="font-family: Arial,sans-serif; font-size: 10pt;">TCCATGCAGCCGATCAGTGGGCTGTCGAACCCGCGACGGTCGACGACGTCCTCGACGCACAACGTTGGGCCCGAG <span style="font-family: Arial,sans-serif; font-size: 10pt;">AGCGCGCACAGAGAGCAGTTAGCGGTATGGCGAGTGTAGCTATTGCGTCAACGGCAAAGCCGGGCGCTGCCGGTA <span style="font-family: Arial,sans-serif; font-size: 10pt;">GGCACGCCAGCACCCTAGAAAGATCA


 * *GC% Content for gene: 56.7% ( ** [] )
 * *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): **