Serine-threonine+protein+kinase+PknG+(Mycobacterium+leprae)

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Mycobacterium leprae causes an infectious disease called Leprosy (or commonly known as the Hansen's Disease). Leprosy affects the peripheral nerves, the skin, the upper respiratory tract, eyes, and the nasal mucus. Symptoms of Leprosy may show years later. Leprosy survive best at low temperatures and effects of this develops irrationally. Those with poor T-cell immunity towards M. leprae typically exhibit lepromatous (LL) leprosy or multi-bacillary leprosy, which is associated with symmetric skin lesions, nodules, plaques, thickened dermis, and frequent involvement of the nasal mucosa resulting in congestion and nose bleeds. Leprosy may cause some degree of peripheral neurological damage as well as muscle weakness. People with long-term leprosy may lose the use of their hands or feet due to repeated traumatic injury resulting from lack of sensation. If left untreated, it can cause progressive and permanent damage to the skin, nerves, eyes, and limbs.
 * *Target (protein/gene name): **Serine-threonine protein kinase PknG (protein kinase G) (STPK G) ======
 * *NCBI Gene # or RefSeq#: **15827075
 * *Protein ID (NP or XP #) or Wolbachia#: ** [|NP_301338.1]
 * *Organism (including strain): ** Mycobacterium leprae strain TN
 * Etiologic Risk Group (see link below): ****[|NIH RG 2] **
 * */Disease Information (sort of like the Intro to your Mini ****__Research Write __**** up): **


 * Link to TDR Targets page (if present): ****[|click!] **
 * <span style="font-family: Arial,sans-serif;">Link to Gene Database page (__[|NCBI]__, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or [|PATRIC], etc.) **

<span style="color: #4f81bd; font-family: Arial,sans-serif;">S <span style="color: #4f81bd; font-family: Arial,sans-serif;">erine-threonine protein kinase PknG <span style="color: #4f81bd; font-family: Arial,sans-serif;">is known to promote mycobacterial survival in macrophage. It does so by being secreted into the cytosol of infected macrophages and inhibits phagosome-lysosome fusion. The pknG gene is the terminal gene in a putative operon containing glnH, encoding a protein potentially involved in glutamine uptake.
 * <span style="font-family: Arial,sans-serif; line-height: 1.5;">Essentiality of this protein: **

<span style="font-family: Arial,sans-serif; line-height: 1.5;">**Is it a monomer or multimer as biological unit? (make prediction at** @http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html): <span style="color: #4f81bd; font-family: Arial,sans-serif; line-height: 1.5;">monomer
 * <span style="font-family: Arial,sans-serif;">Complex of proteins?: **<span style="color: #4f81bd; font-family: Arial,sans-serif;">monomer
 * <span style="font-family: Arial,sans-serif;">Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): **<span style="color: #4f81bd; font-family: Arial,sans-serif;">corynebactieral PknG (CgPknG) - homology [|(click link!)]


 * <span style="font-family: Arial,sans-serif;">*EC#: ****<span style="color: #4f81bd; font-family: Arial,sans-serif;">2.7.11.1 **


 * <span style="font-family: Arial,sans-serif;">Link to BRENDA EC# page: ** [|click!]
 * <span style="font-family: Arial,sans-serif;">-- **<span style="font-family: Arial,sans-serif;"> Show [|screenshot] of BRENDA enzyme mechanism schematic

Figure 1: Reaction catalyzed by serine/threonine protein kinase (2.7.11.1) from BRENDA.
<span style="font-family: Arial,sans-serif;"> [] <span style="color: #4f81bd; font-family: Arial,sans-serif;">Been discontinued- CS0730 but costs $766.00
 * <span style="font-family: Arial,sans-serif;">Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): **
 * <span style="font-family: Arial,sans-serif;">-- link to Sigma (or other company) page for assay (see Sigma links below) **
 * <span style="font-family: Arial,sans-serif;">---or link (or citation) to paper that contains assay information **[|**Kinase assays**]
 * <span style="font-family: Arial,sans-serif;">-- links to assay reagents (substrates) pages. ****<span style="color: #4f81bd; font-family: Arial,sans-serif; line-height: 1.5;">[|See paper] **
 * <span style="font-family: Arial,sans-serif;">--- List cost and quantity of substrate reagents, supplier, and catalog # **


 * <span style="font-family: Arial,sans-serif;">Structure (PDB or Homology model) **

<span style="font-family: Arial,sans-serif; line-height: 0px; overflow: hidden;">

Figure 2. PDB Crystal Structure of Protein Kinase PknG (4Y0X)

<span style="font-family: Arial,sans-serif;">-- PDB # or closest PDB entry if using homology model: <span style="color: #4f81bd; font-family: Arial,sans-serif;">[|4Y0X]
 * <span style="font-family: Arial,sans-serif;">Current Inhibitors: ** <span style="color: #4f81bd; font-family: Arial,sans-serif;">[|K-252a, staurosporine, KbpA]
 * <span style="font-family: Arial,sans-serif; line-height: 1.5;">Expression Information (has it been expressed in bacterial cells): **<span style="color: #4f81bd; font-family: Arial,sans-serif; line-height: 1.5;">E. coli BL21 (DE3)
 * <span style="font-family: Arial,sans-serif;">Purification Method: **<span style="color: #4f81bd; font-family: Arial,sans-serif; font-size: 10pt;">His-tag Ni-NTA affinity purification


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

Figure 3. Pymol Structure of Protein Kinase PknG(4Y0X)shown as sticks. Carbons are colored as cyan and surface transparency set to 80%

 * <span style="font-family: Arial,sans-serif;">*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): **

==**<span style="font-family: Arial,sans-serif; font-size: 13px; line-height: 1.5;">Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website ****<span style="color: #4f81bd; font-family: Arial,sans-serif; font-size: 13px; line-height: 1.5;">83888.2 kDa **== ==**<span style="font-family: Arial,sans-serif; font-size: 13px; line-height: 1.5;">Molar Extinction coefficient of your protein at 280 nm wavelength: ****<span style="color: #4f81bd; font-family: Arial,sans-serif; font-size: 13px; line-height: 1.5;">67310 **== ==**<span style="font-family: Arial,sans-serif; font-size: 13px; line-height: 1.5;">TMpred graph Image **<span style="font-family: Arial,sans-serif; font-size: 13px; line-height: 1.5;"> (@http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it. ==
 * <span style="font-family: Arial,sans-serif;">*length of your protein in Amino Acids ****<span style="color: #4f81bd; font-family: Arial,sans-serif;">787 amino acids **

[[image:graph- virtual discovery.JPG width="613" height="477"]]
<span style="background-color: #ffffff; color: #4f81bd; font-family: 'Courier New'; font-size: 10pt; line-height: 1.5;">**Figure 4. TMpred output from the amino acid sequence of Protein Kinase PknG(4Y0X).**

<span style="color: #4f81bd; font-family: Arial,sans-serif; font-size: 10pt;">1 maesmkrehm dhdtedvgqa aqradppsgt tegrlqstqa ifrpnfdddd dllhisvpsv <span style="color: #4f81bd; font-family: Arial,sans-serif; font-size: 10pt;">61 dtepqdritp atrvlppirq lggglveirr vrdidpleal mtnpvvpesk rfcwncgrpv <span style="color: #4f81bd; font-family: Arial,sans-serif; font-size: 10pt;">121 grselegqea dgaqgakegw cpycgspysf lpqlspgdiv agqyevkgci ahgglgwvyl <span style="color: #4f81bd; font-family: Arial,sans-serif; font-size: 10pt;">181 afdhnvndrp vvlkglvhsg daeaqasava erqflaevvh pqivqifnfv ehkdtsgdpv <span style="color: #4f81bd; font-family: Arial,sans-serif; font-size: 10pt;">241 gyivmeyigg rslkrgskkg nveklpvaea iaylleilpa lsylhsiglv yndlkpenim <span style="color: #4f81bd; font-family: Arial,sans-serif; font-size: 10pt;">301 lteeqlklid lgavsrinsf gciygtpgyq apeivrtgpt vatdiytvgr tlaaltlnlr <span style="color: #4f81bd; font-family: Arial,sans-serif; font-size: 10pt;">361 trngrymdgl peddpvltty dsfarllhra inpdprrrfs saeemsaqlm gvlrevvaqd <span style="color: #4f81bd; font-family: Arial,sans-serif; font-size: 10pt;">421 tgvpraglst ifspsrstfg vdllvahtdv yldgrlhsek ltakdivtal qvplvdptdv <span style="color: #4f81bd; font-family: Arial,sans-serif; font-size: 10pt;">481 aapvlqatvl sqpvqtldsl raarhgmlda qgidlaesve lplmevrall dlgdvvkanr <span style="color: #4f81bd; font-family: Arial,sans-serif; font-size: 10pt;">541 klddladrvs cqwrlvwyra vadlltgdya satkhftevl ntfpgelapk lalaataela <span style="color: #4f81bd; font-family: Arial,sans-serif; font-size: 10pt;">601 gesdehkfyr tvwhtndgvv saafglarfq saegdrtgav ctldevppts rhfttarlts <span style="color: #4f81bd; font-family: Arial,sans-serif; font-size: 10pt;">661 avtllsgrst neiteqqird aarrvetlpp teprvlqira lvlgcamdwl adnqasanhi <span style="color: #4f81bd; font-family: Arial,sans-serif; font-size: 10pt;">721 lgfpftkhgl rlgveaslrs larvaptqrh rytlvdmank vrptstl
 * <span style="font-family: Arial,sans-serif;">*CDS Gene Sequence (paste as text only): **


 * <span style="font-family: Arial,sans-serif;">*GC% Content for gene: ** **<span style="color: #4f81bd; font-family: Arial,sans-serif;">8.452536% **
 * <span style="font-family: Arial,sans-serif;">*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only): **
 * <span style="font-family: Arial,sans-serif;">*GC% Content for gene (codon optimized): **

<span style="font-family: Arial,sans-serif;">Do Not Need this info for Spring (but still copy these lines to your Target page for now) <span style="font-family: Arial,sans-serif;">-- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to
 * <span style="font-family: Arial,sans-serif;">Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): **
 * <span style="font-family: Arial,sans-serif;">( ****<span style="color: #008000; font-family: Arial,sans-serif;">link to DNA Works output text file - **<span style="font-family: Arial,sans-serif;">that should be saved in your Google Docs folder after you did the primer design protocol)
 * <span style="font-family: Arial,sans-serif;">Primer design results for 'tail' primers (this is just 2 sequences): **