Carbamate+Kinase, Giardia+Lamblia


 * *Target (protein/gene name): ** Carbamate Kinase

GL50803_16453
 * *NCBI Gene # or RefSeq#: **

NW_002477112.1
 * *Protein ID (NP or XP #) or Wolbachia#: **


 * *Organism (including strain): ** Giardia Lamblia

Group 1 http://www.bacterio.net/-hazard.html#group1
 * Etiologic Risk Group (see link below): **

Giardiasis is a common intestinal infection caused by the exposure to the organism Giardia lamblia [1]. Exposure to contaminated water, food, or direct contact with the organism can lead to the disease [1]. Giardia is one of the most common intestinal parasites in the world making Giardiasis a very common infection. Cases are becoming increasingly common in the United States. A well-known cause of Giardiasis is through drinking unfiltered water that may be contaminated with human or animal waste [1]. This has brought the infectious disease to be highly associated with nicknames “travelers diarrhea” and “beaver fever”. One of the main concerns is that children can easily spread the infection in a daycare environment. The increased contact between children and each other’s feces makes it a likely spreading ground [1]. When the inactive forms of Giardia (Cysts) are ingested, the acid within the stomach releases the parasite from the cyst. From there the parasite latches to the lining of the small intestine and reproduces. Cysts are then passed on through the fecal matter of the host [1]. Giardiasis is non fatal and a large number of those infected never display symptoms. When symptoms are displayed they often involve watery and aggressive diarrhea other symptoms include stomach pain, nausea, and vomiting. The disease can often be allowed to run its course or treated with a multitude of antibiotics.
 * */ Disease Information (sort of like the Intro to your Mini Research Write up): **

[1] Carol A. Turkington, Giardiasis, The Gale Encyclopedia of Medicine, 2011, vol. 3, (4th ed.), 1889-1892 https://www.ncbi.nlm.nih.gov/gene/5701142
 * Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.) **

The Carbamate Kinase is essential to the function of Giardia Lamblia as is is part of a pathway that contributes to energy production. Carbamate Kinase catalyzes the reversible conversion of carbamoyl phosphate and ADP to ATP, ammonia and CO 2
 * Essentiality of this protein: **


 * Is it a monomer or multimer as biological unit? ** Multimer as a biological unit

http://www.rcsb.org/structure/4OLC
 * Complex of proteins?: ** yes

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3565245/ (inhibition in Giardia Lamblia)
 * Druggable Target : **


 * *EC#: ** 2.7.2.2

https://www.brenda-enzymes.org/enzyme.php?ecno=2.7.2.2
 * Link to BRENDA EC# page: **
 * -- ** Show screenshot of BRENDA enzyme mechanism schematic

ADP + carbamoyl phosphate
 * Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): **

ATP + carbamate

ATP + NH3 + CO2

https://www.sigmaaldrich.com/catalog/product/supelco/46856u?lang=en®ion=US https://www.sigmaaldrich.com/catalog/product/sigma/flaas?lang=en®ion=US
 * -- link to Sigma (or other company ) page for assay (see Sigma links below) **

-- PDB # or closest PDB entry if using homology model: **4OLC**
 * Structure (PDB or Homology model) **

https://www.ncbi.nlm.nih.gov/pubmed/24558036
 * Current Inhibitors: Disulfarim **


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

**Yes, Previously Expressed in E.Coli Cells (BL21)** DE3)Star as a TEV protease-cleavable, His-tagged, maltose binding protein fusion protein
possible method: https://www.sciencedirect.com/science/article/pii/0926656964900100
 * Purification Method : **


 * Image of protein (PyMol with features delineated and shown separately): **
 * *Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): **

=
 MSAGKTVVIA LGGNAMLQAK EKGDYDTQRK NVEIAASEIY KIHKAGYKVV LTSGNAPQVG AIKLQNQAAA GVSPEMPLHV CGAMSQGFIG YMMSQAMDNV FCANNEPANC VTCVTQTLVD PKDQAFTNPT KPRWRFYTEQ EAKDLMAANP GKILREDAWP AGWRVVVPSP RPLEIVEYGV IKTLIDNNVL VICTNGGGIP CKRENKVISG VDAVIDKDLA TSLLAKTLNS DYLMILTDVL NACINYKKPD ERKLEEIKLS EILALEKDGH FAAGSMGPKV RAAIEFTQAT GKMSIITSLS TAVDALNGKC GTRIIKD =====


 * *length of your protein in Amino Acids: 317 **


 * Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website: 34.220 **


 * Molar Extinction coefficient of your protein at 280 nm wavelength: ****Ext. coefficient 28420 Abs 0.1% (=1 g/l) 0.830, assuming all Cys residues are reduced**


 * 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): **

GGACCTGCTAACCCTTGCTGTGGTGTCTACAGGAGTCACTGCGGGGGCGCTTTCCTCCATAACACTTGTA

GAGTAAAGAATGATCTAAAGTTTTTGATCGTTAAAAGCCTCAGACGCCCTTGAGGCGGAAAATAAATTCC

AGATGTCGGCAGGGAAAACGGTTGTGATTGCGCTGGGCGGCAACGCTATGCTTCAGGCCAAGGAGAAGGG

GGACTATGACACTCAACGTAAGAATGTTGAGATTGCGGCCTCGGAGATTTATAAAATCCATAAGGCTGGC

TATAAGGTTGTTCTCACAAGTGGTAACGGGCCGCAGGTGGGTGCCATAAAACTCCAAAACCAGGCTGCAG

CAGGTGTTTCTCCAGAAATGCCCCTCCACGTCTGTGGTGCAATGTCCCAGGGATTCATCGGCTACATGAT

GTCTCAGGCGATGGACAACGTTTTTTGCGCGAATAACGAGCCAGCAAACTGTGTTACCTGTGTCACTCAG

ACATTAGTCGATCCGAAGGATCAGGCTTTCACAAATCCCACAAAGCCCGTTGGGAGGTTCTACACCGAGC

AGGAGGCGAAGGACCTCATGGCTGCAAACCCGGGAAAAATCCTCCGCGAGGATGCTGGCCGCGGCTGGCG

TGTTGTCGTCCCATCTCCTCGTCCTCTTGAGATCGTGGAGTATGGTGTCATCAAGACCCTCATTGACAAC

AACGTTCTTGTCATTTGTACGAATGGGGGAGGAATCCCTTGCAAGCGCGAGAACAAGGTCATCAGTGGCG

TGGATGCTGTCATCGACAAGGACCTTGCAACCTCTCTTCTAGCTAAAACGCTGAACTCGGACTACCTTAT

GATACTTACGGACGTCCTCAACGCGTGCATCAATTACAAGAAGCCCGATGAGAGAAAGCTCGAGGAGATC

AAGCTTAGCGAGATTCTCGCCCTCGAAAAGGACGGCCACTTTGCTGCCGGATCTATGGGGCCGAAGGTCC

GTGCAGCAATCGAGTTTACACAGGCAACTGGAAAAATGAGCATCATCACCTCCCTGAGCACTGCGGTGGA

TGCGCTCAATGGTAAATGTGGGACCCGCATCATCAAGGATTAACTTTGAAGCGAGGCTGTACGCTCCCAC

TCATATATCAGTGAATTTTCCAGCTGCTCTACCTTCGTCATTAGCCGCATAATCTCGTCCTCGTATGGCT

TGCCTGCCCTGTTGAGCCGAAGAGCAGTATTCAGCTGTTCGATCTCT


 * *GC% Content for gene: ** 51.4%


 * *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)