D-alanine+ligase+(Yersinia+pestis)

http://www.sigmaaldrich.com/catalog/product/sigma/a7377?lang=en®ion=US **Substrate Quantity:** 5G http://vdsstream.wikispaces.com/file/view/KovacDalanineDalanineLigaseVirtualScreenJMedChem2008.pdf/261437724/KovacDalanineDalanineLigaseVirtualScreenJMedChem2008.pdf code MAEKVAVLLGGTSAEREVSLLSGQAVLAGLKEAGIDAYGVDTKDFPVTQLKEQGFDKVFI ALHGRGGEDGTLQGVLEFLQLPYTGSGVMASALTMDKLRTKLVWQALGLPISPYVALNRQ QFETLSPEELVACVAKLGLPLIVKPSHEGSSVGMSKVDHASELQKALVEAFQHDSDVLIE KWLSGPEFTVAILGDEVLPSIRIQPPGVFYDYDAKYLSDKTQYFCPSGLSDESEQQLAAL ALQAYHALDCSGWGRVDVMQDRDGHFYLLEVNTSPGMTSHSLVPMAARQYGLSFSQLVAR ILMLADX code code >NC_003143.1:602769-603689 Yersinia pestis CO92 chromosome, complete genome ATGGCTGAGAAAGTCGCGGTATTGCTGGGTGGGACCTCTGCGGAGCGTGAAGTGTCATTACTGTCAGGTC AGGCGGTATTGGCAGGCTTGAAAGAAGCCGGTATCGATGCTTATGGGGTGGATACCAAAGATTTTCCGGT AACTCAGCTTAAAGAACAGGGCTTTGATAAAGTGTTTATCGCCCTGCATGGTCGTGGTGGCGAAGACGGT ACGTTACAAGGCGTATTGGAATTTCTACAACTGCCATATACCGGCAGTGGTGTAATGGCATCCGCCCTGA CAATGGACAAATTGCGCACCAAATTAGTCTGGCAAGCCTTGGGGTTGCCTATTTCGCCTTATGTTGCGTT GAATCGCCAACAGTTTGAAACACTTTCCCCAGAGGAATTGGTGGCGTGTGTCGCTAAGCTTGGCCTGCCA CTTATTGTCAAGCCAAGCCATGAAGGCTCTAGCGTTGGGATGAGTAAAGTTGATCACGCCAGCGAATTGC AGAAGGCATTGGTTGAAGCTTTCCAACATGACAGCGATGTACTGATTGAGAAATGGCTGAGCGGGCCAGA GTTTACTGTCGCGATCCTTGGGGATGAAGTTTTGCCTTCTATTCGTATTCAGCCTCCGGGCGTGTTTTAC GATTATGACGCTAAGTATTTGTCAGATAAAACACAATACTTCTGCCCAAGCGGTTTAAGTGATGAGTCGG AGCAACAATTGGCAGCGCTGGCATTGCAGGCTTACCACGCGCTGGATTGCAGCGGTTGGGGGCGGGTCGA TGTGATGCAAGACCGCGATGGTCATTTTTACCTGCTTGAAGTGAACACTTCACCGGGTATGACCAGCCAT AGTTTGGTTCCAATGGCCGCCCGCCAGTATGGGTTGAGTTTCTCCCAATTGGTGGCGCGCATTCTGATGT TGGCTGACTGA code code format="a-b-r-La"  1 ATGGCGGAAAAAGTTGCGGTTCTC  24 2 ACAGCAGAGAAACTTCACGTTCCGCAGAGGTACCACCCAGGAGAACCGCAACTTTTTCCG 60 3 AACGTGAAGTTTCTCTGCTGTCTGGTCAGGCAGTTCTGGCGGGTCTGAAAGAAGCTGGCA 60 4 GGTAACCGGGAAGTCTTTGGTGTCAACACCGTACGCGTCGATGCCAGCTTCTTTCAGACC 60 5 CCAAAGACTTCCCGGTTACCCAGCTGAAGGAACAGGGTTTCGACAAAGTTTTCATCGCTC 60 6 CGCCCTGGAGGGTACCGTCTTCACCACCACGACCGTGCAGAGCGATGAAAACTTTGTCGA 60 7 GGTACCCTCCAGGGCGTTCTGGAGTTCCTGCAGCTCCCGTACACCGGTTCTGGTGTTATG 60 8 CAGTTTGGTGCGCAGTTTGTCCATGGTCAGTGCGCTCGCCATAACACCAGAACCGGTGTA 60 9 CAAACTGCGCACCAAACTGGTTTGGCAGGCGCTGGGCCTGCCGATCTCTCCATACGTTGC 60 10 TTCTTCCGGAGACAGCGTTTCGAACTGCTGACGGTTCAGCGCAACGTATGGAGAGATCGG 60 11 ACGCTGTCTCCGGAAGAACTGGTAGCGTGCGTTGCGAAACTGGGTCTGCCGCTGATCGTG 60 12 CAACTTTAGACATACCAACAGAAGAACCTTCGTGAGACGGTTTCACGATCAGCGGCAGAC 60 13 TCTTCTGTTGGTATGTCTAAAGTTGACCACGCCTCTGAACTGCAAAAAGCGCTGGTTGAA 60 14 AGCCATTTTTCGATCAGAACGTCAGAGTCGTGCTGGAACGCTTCAACCAGCGCTTTTTGC 60 15 CGTTCTGATCGAAAAATGGCTGAGCGGTCCGGAATTCACCGTTGCTATCCTGGGCGACGA 60 16 GTAGAAAACACCTGGCGGCTGGATACGGATAGACGGCAGAACTTCGTCGCCCAGGATAGC 60 17 GCCGCCAGGTGTTTTCTACGACTACGACGCGAAATACCTGTCTGACAAGACGCAATACTT 60 18 GCTGCTGTTCAGATTCATCGGACAGGCCAGACGGGCAGAAGTATTGCGTCTTGTCAGACA 60 19 CGATGAATCTGAACAGCAGCTCGCGGCACTGGCGCTGCAAGCGTACCACGCGCTGGACTG 60 20 ACCGTCACGGTCCTGCATAACGTCAACGCGACCCCAACCAGAGCAGTCCAGCGCGTGGTA 60 21 GCAGGACCGTGACGGTCACTTCTACCTGCTCGAAGTTAACACGAGCCCGGGTATGACCTC 60 22 AGACAGACCGTACTGACGTGCTGCCATCGGTACCAGAGAGTGAGAGGTCATACCCGGGCT 60 23 ACGTCAGTACGGTCTGTCTTTCTCTCAGCTCGTAGCGCGTATCCTGATGCTGGCGGACTA 60 24 TTAGTCCGCCAGCATCAGG 19 code YpDAL_for: TACTTCCAATCCATGGCGGAAAAAGTTGCGGTTCT YpDAL_rev: TATCCACCTTTACTGTTAGTCCGCCAGCATCAGGA
 * Target: ** D-alanine D-alanine ligase
 * NCBI Gene # or RefSeq#: ** NC_003143.1
 * Protein ID or Wolbachia#: ** YP_002345628.1
 * Organism: **// Yersinia pestis //
 * Etiologic Risk Group: ** Risk Group 3
 * Disease Information : ** //Yersinia pestis// is a bacterium that causes a variation of plague in humans and other mammals. Humans typically get plague after being bitten by a rodent flea that is carrying the plague bacterium, encountering contaminated fluids or tissue, or inhaling infections droplets. Plague is notorious for killing millions of people in Europe during the Middle Ages. Since that time, plague has occurred in rural and semi-rural areas of the western United States. Without immediate treatment, the disease can cause serious illness or death. The most common form of plague is bubonic plague, which infects the lymphatic system and causes inflammation. When the bacteria enter the bloodstream, the disease is then recognized as septicemic plague. Alternatively, if the bacteria spread to the lungs, the disease is then considered to be pneumonic plague, which is the only form of plague that can be transmitted between people. One of the enzymes that helps //Yersinia pestis// infect its host is D-alanine D-alanine ligase, which synthesizes D-alanyl-D-alanine, an essential precursor of bacterial peptidoglycan. For this reason, D-alanine ligase is an important target for the development of antibacterial drugs.
 * Link to TDR Targets page: ** N/A
 * Link to Gene Database page: ** https://www.ncbi.nlm.nih.gov/gene/1173401
 * Essentiality of this Protein: ** D-alanine D-alanine ligase is an essential enzyme in bacterial cell wall biosynthesis.
 * Is it a monomer or multimer as biological unit? ** Multimer
 * Complex of Proteins? ** D-alanine D-alanine ligase consists of three domains, in which four loops, loop 1, loop 2, loop 3 and loop 4, constitute the binding sites for two D-alanine molecules and one ATP molecule.
 * Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): ** https://www.ncbi.nlm.nih.gov/pubmed/23286234
 * EC#: ** 6.3.2.4
 * Link to BRENDA EC# page: ** https://www.brenda-enzymes.org/enzyme.php?ecno=6.3.2.4&Suchword=&reference=&UniProtAcc=&organism%5B%5D=Yersinia+pestis&show_tm=0
 * Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): ** http://vdsstream.wikispaces.com/file/view/KovacDalanineDalanineLigaseVirtualScreenJMedChem2008.pdf/261437724/KovacDalanineDalanineLigaseVirtualScreenJMedChem2008.pdf
 * Link to Sigma page for Assay: ** http://scripts.iucr.org/cgi-bin/paper?S2059798315021671
 * Substrate Cost: ** $37.40
 * Substrate Catalog #: ** A7377-5G
 * PDB Structure: ** 5C1P
 * Current Inhibitors: **
 * Expression Information (has it been expressed in bacterial cells): ** D-alanine D-alanine ligase has been expressed in //E. coli//.
 * Purification Method : ** Ni-NTA
 * Image of Protein: **
 * Amino Acid Sequence: **
 * Length of your protein in Amino Acids: ** 307 AA
 * Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website: ** 33261.23
 * Molar Extinction coefficient of your protein at 280 nm wavelength: ** 31525
 * TMpred graph Image ** (@http://www.ch.embnet.org/software/TMPRED_form.html):
 * CDS Gene Sequence : **
 * GC% Content for Gene: ** N/A
 * CDS Gene Sequence (codon optimized): **
 * GC% Content for Gene (codon optimized): ** N/A
 * Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): **
 * DNA Works File:** https://drive.google.com/drive/folders/0B4O2KqKh2q_-dFJuSVJtcDgtUUU
 * Primer Design Results for 'Tail' Primers: **