TargetSp17-+ATP-+dependent+Clp+protease+proteolytic+subunit

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 * Resource for potential targets: **


 * Target (protein/gene name): ** ClpP; ATP-dependent Clp protease proteolytic subunit


 * *NCBI Gene # or RefSeq#: **3192361


 * *Protein ID (NP or XP #) or Wolbachia#: [|393115] **


 * *Organism (including strain): **// Francisella tularensis //


 * Etiologic Risk Group (see link below): [|Category A Priority (NIAID)] **

Tularemia is a highly contagious disease caused by the bacterium //Francisella tularensiser// that causes fever, formation of ulceration, and sometimes severe pneumonia. There are several types of infection, including glandular, oropharyngeal, pneumonic, oculoglandular, typhoidal, and the most common type, ulceroglandular. The mortality rate isn't high (<1% if treated), but there is high potential for use in bioterrorism because the bacterium is easy to spread, is highly infective, and is highly deblilitative. Most cases reported result from contact with other infected mammals, deer flies, or ticks, and the CDC reports 0.1 cases per 100,000 residents in the US in 2016 ([|CDC]). The clpP gene in //F. tularensis// is responsible for stress tolerance and infection in a mammalian host. clpP is a protease (EC 3.4.21.92) that helps maintain protein homeostasis in the bacterium through a wide range of functions.
 * * Disease Information : **


 * Link to TDR Targets page (if present): [|Here] **


 * Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.)[| Here] **


 * Essentiality of this protein: ** In //M. tuberculosis//, inhibition and activation of the Clp series had the potential to kill the cell ([|source])

Is it a monomer or multimer as biological unit ** ? (make prediction at ** @http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html): Multimer


 * Complex of proteins?: - is this functional as a monomer?? - DR. B **

@http://jb.asm.org/content/194/3/663.full []
 * Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): **

This is a very generalized formula because clpP is a protease with many substrates.
 * *EC#: 3.4.21.92 **
 * [|BRENDA] **

[|Assay Information]
 * Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): **

-- PDB # or closest PDB entry if using homology model: [|5G1Q], [|5G1R] Show pairwise alignment of your BLASTP search in NCBI against the PDB
 * Structure (PDB or Homology model) **

Query Coverage: 1st chain Max % Identities: 56% % Positives: 75% Chain used for homology:

>5G1Q:A|PDBID|CHAIN|SEQUENCE

MITNNLVPTVIEKTAGGERAFDIYSRLLKERIVFLNGEVNDHSANLVIAQLLFLESEDPDKDIYFYINSPGGMVTAGMGVYDTMQFIKPDVSTICIGLAASMGSLLLAGGAKGKRYSLPSSQIMIHQPLGGFRGQASDIEIHAKNILRIKDRLNKVLAHHTGQDLETIVKDTDRDNFMMADEAKAYGLIDHVIESREAIIK


 * Current Inhibitors: ** Beta-lactone D3, Beta-lactone U1, Beta-lactone 7, Phenyl Ester AV170
 * Expression Information (has it been expressed in bacterial cells): ** Expressed in E. coli Bl21 (DE3)
 * Purification Method : ** Ni affinity Chromatography
 * Image of protein (PyMol with features delineated and shown separately): **
 * *Amino Acid Sequence: **

>5G1Q:A|PDBID|CHAIN|SEQUENCE MITNNLVPTVIEKTAGGERAFDIYSRLLKERIVFLNGEVNDHSANLVIAQLLFLESEDPDKDIYFYINSPGGMVTAGMGVYDTMQFIKPDVSTICIGLAASMGSLLLAGGAKGKRYSLPSSQIMIHQPLGGFRGQASDIEIHAKNILRIKDRLNKVLAHHTGQDLETIVKDTDRDNFMMADEAKAYGLIDHVIESREAIIK

>5G1Q:B|PDBID|CHAIN|SEQUENCE MITNNLVPTVIEKTAGGERAFDIYSRLLKERIVFLNGEVNDHSANLVIAQLLFLESEDPDKDIYFYINSPGGMVTAGMGVYDTMQFIKPDVSTICIGLAASMGSLLLAGGAKGKRYSLPSSQIMIHQPLGGFRGQASDIEIHAKNILRIKDRLNKVLAHHTGQDLETIVKDTDRDNFMMADEAKAYGLIDHVIESREAIIK

>5G1Q:C|PDBID|CHAIN|SEQUENCE MITNNLVPTVIEKTAGGERAFDIYSRLLKERIVFLNGEVNDHSANLVIAQLLFLESEDPDKDIYFYINSPGGMVTAGMGVYDTMQFIKPDVSTICIGLAASMGSLLLAGGAKGKRYSLPSSQIMIHQPLGGFRGQASDIEIHAKNILRIKDRLNKVLAHHTGQDLETIVKDTDRDNFMMADEAKAYGLIDHVIESREAIIK

>5G1Q:D|PDBID|CHAIN|SEQUENCE MITNNLVPTVIEKTAGGERAFDIYSRLLKERIVFLNGEVNDHSANLVIAQLLFLESEDPDKDIYFYINSPGGMVTAGMGVYDTMQFIKPDVSTICIGLAASMGSLLLAGGAKGKRYSLPSSQIMIHQPLGGFRGQASDIEIHAKNILRIKDRLNKVLAHHTGQDLETIVKDTDRDNFMMADEAKAYGLIDHVIESREAIIK

>5G1Q:E|PDBID|CHAIN|SEQUENCE MITNNLVPTVIEKTAGGERAFDIYSRLLKERIVFLNGEVNDHSANLVIAQLLFLESEDPDKDIYFYINSPGGMVTAGMGVYDTMQFIKPDVSTICIGLAASMGSLLLAGGAKGKRYSLPSSQIMIHQPLGGFRGQASDIEIHAKNILRIKDRLNKVLAHHTGQDLETIVKDTDRDNFMMADEAKAYGLIDHVIESREAIIK

>5G1Q:F|PDBID|CHAIN|SEQUENCE MITNNLVPTVIEKTAGGERAFDIYSRLLKERIVFLNGEVNDHSANLVIAQLLFLESEDPDKDIYFYINSPGGMVTAGMGVYDTMQFIKPDVSTICIGLAASMGSLLLAGGAKGKRYSLPSSQIMIHQPLGGFRGQASDIEIHAKNILRIKDRLNKVLAHHTGQDLETIVKDTDRDNFMMADEAKAYGLIDHVIESREAIIK

>5G1Q:G|PDBID|CHAIN|SEQUENCE MITNNLVPTVIEKTAGGERAFDIYSRLLKERIVFLNGEVNDHSANLVIAQLLFLESEDPDKDIYFYINSPGGMVTAGMGVYDTMQFIKPDVSTICIGLAASMGSLLLAGGAKGKRYSLPSSQIMIHQPLGGFRGQASDIEIHAKNILRIKDRLNKVLAHHTGQDLETIVKDTDRDNFMMADEAKAYGLIDHVIESREAIIK


 * *length of your protein in Amino Acids: ** 1428 residues - Just show for 1 chain - Dr. B
 * Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website: ** 154.95 kDa Just show for 1 chain - Dr. B


 * Molar Extinction coefficient of your protein at 280 nm wavelength: ** 62955 M-1 cm-1
 * TMpred graph Image ** (@http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it. - Just show for 1 chain - Dr. B

ATGTGGCCCAGAGTGCTGCTGGGGGAGGCCCGGGTGGCTGTGGACGGATGTCGCGCTCTGTTGTCTCGCC
 * *CDS Gene Sequence (paste as text only): **

TTGCCGTGCATTTCTCCCCGCCATGGACTGCTGTGAGCTGCTCACCCCTGCGGAGGAGCCTGCATGGAAC

TGCGACGCGAGCTTTCCCGCTCATCCCCATAGTGGTGGAGCAGACGGGTCGAGGCGAGCGCGCTTATGAC

ATATACTCGAGGCTGTTGCGGGAACGCATCGTGTGCGTCATGGGCCCGATTGACGACAGTGTGGCCAGTC

TGGTCATTGCCCAGCTGTTGTTCTTACAGTCTGAAAGCAACAAGAAGCCCATTCATATGTATATCAACAG

CCCAGGTGGTGTGGTAACTGCGGGCCTGGCCATCTACGACACAATGCAGTACATCCTGAACCCCATCTGC

ACGTGGTGTGTTGGACAGGCTGCCAGCATGGGCTCCCTGCTCCTCGCTGCTGGCAGCCCGGGCATGCGCC

ATTCACTGCCCAATTCCAGAATCATGATCCACCAGCCCTCTGGAGGAGCCAGGGGCCAAGCCACAGACAT

CGCCATCCAGGCAGAGGAAATCATGAAGCTGAAAAAGCAGCTATACAACATCTACGCCAAACACACCAAG

CAGAGCCTACAGGTGATCGAGTCAGCAATGGAGAGGGACCGCTACATGAGCCCCATGGAGGCCCAAGAGT

TTGGCATCTTGGACAAGGTCTTGGTCCACCCACCTCAGGACGGGGAGGATGAGCCAGAACTGGTACAGAA

GGAGACTGCCACAGCGCCGACGGATCCTCCTGCCCCGACAAGCACCTAA
 * *GC% Content for gene: ** 58.73%

Do Not Need this info for Spring (but still copy these lines to your Target page for now)


 * output**


 * *GC% Content for gene (codon optimized): **

-- 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)

** Links:
 * Primer design results for 'tail' primers (this is just 2 sequences): **

[|Stress Tolerance and Infection NCBI] [|Open and Closed Conformations]