GuaA+(Borrelia+burgodorferi)


 * *Target (protein/gene name): GuaA **

***NCBI Gene # or RefSeq#:** NCBI Gene # 224308

***Protein ID (NP or XP #) or Wolbachia#:** NP #047004.2

***Organism (including strain):** Borrelia burgdoferi

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


 * */Disease Information: **

Borrelia burgdorferi is one of the four bacteria species that causes Lyme Disease, a vector-born—usually transmitted by ticks—that starts as a rash, but can develop into chronic arthritis and neurological issues. Unlike most bacteria, Borrelia burgorferi has two membranes with peptidoglycan between them, but the outer membrane does not have lipopolysaccharides. This allows Borrelia burgdorferi to evade its host’s immune response; moreover, Borrelia burgdorferi can change its plasma membrane glycoproteins and proteases to prevent immune response. GuaA—the gene that encodes for GMP synthase—is needed for Borrelia burgdorferi’s survival during the infection cycle. GuaA promotes Borrelia burgdorferi’s growth by synthesizing GMP from XMP, a vital process of purine metabolism.

**Link to TDR Targets page (if present):** N/A

**Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.):** http://www.ncbi.nlm.nih.gov/protein/NP_047004.2

**Essentiality of this protein:** GuaA is essential for Borrelia burgdorferi survival in mice. It encodes for GMP synthase, which is needed to form purine bases in DNA.

**Complex of proteins?:** No.

**Druggable Target:** Amoxicillin-clavulanic acid, ceftriaxone, and high-dosage penicillin G are known inhibitors, while oxytetracycline, doxycycline, chloramphenicol, erythromycin, and azithromycin are known to not cure Borrelia burgdorferi. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC284594/

***EC#:** 6.3.5.2

**Link to BRENDA EC# page:** http://www.brenda-enzymes.org/enzyme.php?ecno=6.3.5.2  <span style="font-family: Arial,Helvetica,sans-serif;">Figure 1) Reaction mechanism detailing the conversion of GMP to XMP with the use of catalyst GuaA.

<span style="font-family: Arial,Helvetica,sans-serif;">**Enzyme Assay information:** [] <span style="font-family: Arial,Helvetica,sans-serif;">Xanthine/Hypoxanthine Assay Kit from Sigma-Aldrich costs $303.50, but includes all of the necessary reagents. Product numbers were not given.
 * <span style="font-family: Arial,Helvetica,sans-serif;">Xanthine assay buffer
 * <span style="font-family: Arial,Helvetica,sans-serif;">Peroxidase substrate
 * <span style="font-family: Arial,Helvetica,sans-serif;">Xanthine enzyme mix
 * <span style="font-family: Arial,Helvetica,sans-serif;">Developer
 * <span style="font-family: Arial,Helvetica,sans-serif;">Xanthine standard

<span style="font-family: Arial,Helvetica,sans-serif;">**Structure (PDB or Homology model):** 1GPM (homology model for E. coli) <span style="font-family: Arial,Helvetica,sans-serif;">

<span style="font-family: Arial,Helvetica,sans-serif;">Figure 2) Complete PyMol structure of the homology model in E. coli (PDB: 1GPM) for the GuaA gene in Borrelia burgorferi. <span style="font-family: Arial,Helvetica,sans-serif;">

<span style="font-family: Arial,Helvetica,sans-serif;">Figure 3) Sequence alignment obtained through BLAST shown for the protein sequence between GuaA in Borrelia burgorferi and Homo sapiens. <span style="font-family: Arial,Helvetica,sans-serif;"> <span style="font-family: Arial,Helvetica,sans-serif;">Figure 4) Sequence alignment obtained through BLAST shown for the protein sequence between GuaA in Borrelia burgorferi and Homo sapiens.

<span style="font-family: Arial,Helvetica,sans-serif;">**Current Inhibitors:** Amoxicillin-clavulanic acid, ceftriaxone, and high-dosage penicillin G are known inhibitors.

<span style="font-family: Arial,Helvetica,sans-serif;">**Expression Information (has it been expressed in bacterial cells):** Yes, GuaA can be expressed in several kinds of bacterial cells, including E. coli.

<span style="font-family: Arial,Helvetica,sans-serif;">**Purification Method:** Buffers PB and PE obtained through the QIAquick PCR Purification Kit (Qiagen, Chatsworth, CA)


 * <span style="font-family: Arial,Helvetica,sans-serif;">Image of protein (PyMol with features delineated and shown separately): **

<span style="font-family: Arial,Helvetica,sans-serif;">MNAQAILVLDFGSQYSQLIARRIREIGVYTKVIPYYTPLKEIKNMNISGIILSGSPASVY <span style="font-family: Arial,Helvetica,sans-serif;">SKEAPTLDMEIFNLKIPVLGICYGMQIIVKLFGGLVSKDSKQEYGRSEIFLKDEKSLLFS <span style="font-family: Arial,Helvetica,sans-serif;">ELPNKFQIIMSHGDSIEKIPDNFKQLAFTKNCIASISNETQKIYGLQFHPEVTHSEFGDQ <span style="font-family: Arial,Helvetica,sans-serif;">ILKNFVFKICQAQINWSLEGNLETIVKKIKLKVGSKKVILGLSGGTDSLVCALLIKKAIN <span style="font-family: Arial,Helvetica,sans-serif;">ENLICVFVNTGLLRKNENKKILELKHQYDLNIKYIDASTKFLNRLKNISDPEEKRKIIGK <span style="font-family: Arial,Helvetica,sans-serif;">EFVDVFEKITLEDQNIEYLAQGTIYSDVIESKSKDSSSSKIKSHHNVGGLPDKMSLKLLE <span style="font-family: Arial,Helvetica,sans-serif;">PLNEFFKDEIIQIGINLGIKKESLYRHPFPGPGLAIRIIGEVTQEKINILQEADNILTEE <span style="font-family: Arial,Helvetica,sans-serif;">LFINDLYYQIRQAFVVLLPVKSVGVMGDQRTYEYTAVIRCVNTQDFMTAEWTELPYSFLK <span style="font-family: Arial,Helvetica,sans-serif;">KVSSRIINEVRGINRVCYDISSKPPSTIEWE
 * <span style="font-family: Arial,Helvetica,sans-serif;">*Amino Acid Sequence: **

<span style="font-family: Arial,Helvetica,sans-serif;">**Length of your protein in Amino Acids:** 511 amino acids

<span style="font-family: Arial,Helvetica,sans-serif;">**Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website:** 58 kDa

<span style="font-family: Arial,Helvetica,sans-serif;">**Molar Extinction coefficient of your protein at 280 nm wavelength:** 45185 1/(M*cm)

<span style="font-family: Arial,Helvetica,sans-serif;">**TMpred graph Image** (@http://www.ch.embnet.org/software/TMPRED_form.html). <span style="font-family: Arial,Helvetica,sans-serif;">
 * <span style="font-family: Arial,Helvetica,sans-serif;">CDS Gene Sequence (paste as text only): **

<span style="font-family: Arial,Helvetica,sans-serif;">atgaacacccaggcgattctggtgctggattttggcagccagtatagccagctgattgcg cgccgcattcgcgaaattggcgtgtataccaaagtgattccgtattataccccgctgaaa gaaattaaaaacatgaacattagcggcattattctgagcggcagcccggcgagcgtgtat agcaaagaagcgccgaccctgaacatggaaatttttaacctgaaaattccgattctgggc atttgctatggcatgcagattattgtgaaactgtttggcggcctggtgagcaaagatagc aaacaggaatatggcagcagcgaaatttttctgcgcgatgaaaaaagcctgctgtttagc gaactgccgaacaaatttcagattattatgagccatggcgatagcattgaaaaaattccg gataactttaaacagctggcgtttaccaaaaactgcattgcgagcattagcaacgaaacc cagaaaatttatggcctgcagtttcatccggaagtgacccatagcgaatttggcgatcag attattaaaaactttgtgtttaaaatttgccaggcgcagattaactggagcctggaaggc aacctggaaaccattgtgaaaaaaattaaactgaaagtgggcagcaaaaaagtgattctg ggcctgagcggcggcaccgatagcctggtgtgcgcgctgctgattaaaaaagcgattaac gaaaacctgatttgcgtgtttgtgaacaccggcctgctgcgcaaaaacgaagataaaaaa attctggaactgaaacatcagtatgatctgaacattaaatatattgatgcgagcaccaaa tttctgaaccgcctgaaaaacattagcgatccggaagaaaaacgcaaaattattggcaaa gaatttgtggatgtgtttgaaaaaattaccctggaagatcagaacattgaatatctggcg cagggcaccatttatagcgatgtgattgaaagcaaaagcaaagatagcagcagcagcaaa attaaaagccatcataacgtgggcggcctgccggataaaatgagcctgaaactgctggaa ccgctgaacgaattttttaaagatgaaattattcagattggcattaacctgggcattaaa aaagaaagcctgtatcgccatccgtttccgggcccgggcctggcgattcgcattattggc gaagtgacccaggaaaaaattaacattctgcaggaagcggataacattctgaccgaagaa ctgtttattaacgatctgtattatcagattcgccaggcgtttgtggtgctgctgccggtg aaaagcgtgggcgtgatgggcgatcagcgcacctatgaatataccgcggtgattcgctgc gtgaacacccaggattttatgaccgcggaatggaccgaactgccgtatagctttctgaaa aaagtgagcagccgcattattaacgaagtgcgcggcattaaccgcgtgtgctatgatatt agcagcaaaccgccgagcaccattgaatgggaa

<span style="font-family: Arial,Helvetica,sans-serif;">***GC% Content for gene:** 44.9%

<span style="font-family: Arial,Helvetica,sans-serif;">*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):

<span style="font-family: Arial,Helvetica,sans-serif;">*GC% Content for gene (codon optimized):