Target+-+Tyrosyl-tRNA+synthetase+(L.+major)


 * *Target (protein/gene name): ** Leishmania major, tyrosyl-tRNA synthetase, putative


 * *NCBI Gene # or RefSeq#: **


 * *Protein ID (NP or XP #) or Wolbachia#: ** LmjF14.1370


 * *Organism (including strain): ** Leishmania major


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


 * *Background/ ** **Disease Information** ** (sort of like the Intro to your Mini Research Write up): ** Leishmania major is a protozoa that causes cutaneous leishmaniasis, which harms the macrophages and dendritic immune cells of the host. It is transmitted by the female sandfly and can cause uncomfortable skin lesions. Leishmaniasis is quickly transmitted from person to person in sandy regions, which are hotspots for sandflies. (The disease may also be transferred through the sharing of needles, but this occurs in only a small minority of cases.) When a sandfly sucks the blood of an infected individual, it takes in the //Leishmania// and becomes a carrier of the disease. Leishmaniasis is transmitted when sandflies carrying the //Leishmania// parasite engage in a subsequent blood meal, injecting the young parasite, or promastigote, into the flesh wound of the human. Macrophages of the host engulf the //Leishmania// promastigote, which matures inside of the host cell and proceeds to infect other macrophages.


 * Link to TDR Targets page (if present): ** http://tdrtargets.org/targets/view?gene_id=27213


 * Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.): ** http://tritrypdb.org/tritrypdb/showRecord.do?name=GeneRecordClasses.GeneRecordClass&primary_key=LmjF14.1370

Gene/Ortholog: Tb927.7.3620 (OG4_11028); Phenotype: significant gain of fitness in procyclic forms; Source study: alsford
 * Essentiality of this protein: **


 * Complex of proteins?: ** yes

Synthetic analogues of SB-219383. Novel C-glycosyl peptides as inhibitors of tyrosyl tRNA synthetase. (2001).
 * Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): **** Homo sapiens (1) **


 * Bioorg Med Chem Lett 11: ** 711-4.


 * *EC#: ** 6.1.1.1


 * Link to BRENDA EC# page: ** http://brenda-enzymes.org/php/result_flat.php4?ecno=6.1.1.1

**Enzyme Assay** ** information (spectrophotometric, coupled assay ?, reagents): **The effect of substrate binding on trypanosomal tyrosyl-tRNA synthetase was assayed by adding substrate or other potential ligands to 0.5 mg/ml protein in standard buffer and monitoring fluorescence over a temperature range of 20–90° C. [] Sigma Aldrich Supplier ATP disodium salt hydrate-$ 37.20 (1mg)-FLAAS L-tyrosine-$65.50 (50g)- T3754 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC390865/pdf/pnas00363-0045.pdf
 * -- ** Show screenshot of BRENDA enzyme mechanism schematic


 * Structure Available (PDB or Homology model) **

-- PDB # or closest PDB entry if using homology model: 3P0J

-- Max % Identities: 100%
 * Show pairwise alignment of your BLASTP search in NCBI against the PDB **

-- % Positives: 100%


 * Current Inhibitors: ** N/A


 * Expression Information (has it been expressed in bacterial cells): ** Yes, expressed in E. coli.

**Purification Method** ** : ** Ni-NTA affinity chromatography


 * Image of protein (PyMol with features delineated and shown separately): **

MNTDERYKLLRSVGEECIQESELRNLIEKKPLIRCYDGFEPSGRMHIAQGIFKAVNVNKC
 * *Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): **

TAAGCEFVFWVADWFALMNDKVGGELEKIRIVGRYLIEVWKAAGMDMDKVLFLWSSEEIT

SHADTYWRMVLDIGRQNTIARIKKCCTIMGKTEGTLTAAQVLYPLMQCCDIFFLKADICQ

LGLDQRKVNMLAREYCDLIGRKLKPVILSHHMLAGLKQGQAKMSKSDPDSAIFMEDTEED

VARKIRQAYCPRVKQSASAITDDGAPVATDDRNPVLDYFQCVVYARPGAAATIDGTTYAT

YEDLEQAFVSDEVSEDALKSCLIDEVNALLEPVRQHFASNEEAHELLEAVKSYRKDGATL

PLAETALPAAPAKPHACMWMPALLKVPLDVAEGMIKVTKDFIAAHPEGTVTLVLPDWSAV

ASDEITGVEKDISAALQVNCALLKAYGLPSSVKIVTENEVILGNCDDFWVSVIGIARKNL

LSHVEELYGGEVRNAGQVIAALMRVATALMLSVSHVISTSLDGHINAFAREYTKERIDCV

QTLEGRIPALHRPGAAPAVLGADDVLYLDDNDMDIRRKIKKAYSAPNEEANPVISVAQHL

LAQHGALSIERGEANGGNVSYNTPEALVADCGSGALHPADLKAAVLQLLLDRSAQARALL

NGELKKNMTVLRNAEKKMAKKR


 * *length of your protein in Amino Acids: ** 682aa

**Molecular Weight** ** of your protein in kiloDaltons using the **[|**Expasy ProtParam**]** website: ** 74.949 kDa


 * Molar ** **Extinction coefficient** ** of your protein at 280 nm wavelength: ** 71945


 * 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): **ATGAACACGGACGAGCGCTACAAGCTCCTCCGCAGCGTCGGAGAGGAGTGCATTCAGGAA

AGTGAGCTGCGCAACCTTATCGAAAAGAAGCCGCTCATTCGCTGCTACGATGGCTTCGAG

CCCTCCGGCCGCATGCACATTGCTCAGGGTATCTTCAAGGCAGTCAATGTCAACAAGTGC

ACCGCCGCCGGGTGCGAGTTTGTCTTCTGGGTGGCGGACTGGTTTGCGCTCATGAACGAC

AAGGTCGGCGGCGAACTGGAGAAGATCCGCATTGTTGGCAGGTACCTTATTGAGGTGTGG

AAGGCGGCGGGCATGGACATGGACAAGGTTTTGTTTTTATGGAGCAGCGAGGAGATCACG

AGCCACGCCGACACGTACTGGCGCATGGTGCTCGACATTGGCCGCCAGAATACGATCGCC

CGCATCAAGAAGTGCTGCACCATCATGGGCAAGACGGAAGGCACGCTGACGGCGGCACAG

GTGCTGTACCCGCTGATGCAGTGCTGCGACATCTTCTTTCTCAAGGCCGACATCTGCCAG

CTGGGCCTGGACCAGCGCAAGGTGAACATGCTCGCACGCGAGTACTGCGATCTGATCGGC

CGCAAGTTGAAGCCGGTCATTCTGTCACACCACATGCTGGCCGGTCTGAAGCAGGGGCAG

GCGAAGATGAGCAAGAGTGACCCGGACAGTGCCATCTTCATGGAAGACACCGAGGAGGAT

GTGGCACGTAAGATCCGTCAGGCCTACTGTCCGCGCGTCAAGCAGTCCGCCAGCGCCATC

ACAGATGACGGGGCGCCGGTCGCCACGGACGACCGCAACCCGGTGCTGGACTACTTCCAG

TGTGTCGTCTACGCCCGACCCGGCGCTGCGGCCACCATTGACGGCACCACCTACGCGACC

TACGAGGACCTCGAGCAGGCTTTTGTGAGCGACGAGGTCAGCGAGGATGCCCTCAAGTCG

TGCCTGATTGATGAGGTGAACGCCCTGCTGGAGCCCGTGCGCCAACATTTTGCGTCAAAC

GAGGAGGCTCACGAGCTGCTGGAGGCGGTGAAGTCGTACCGTAAAGATGGCGCGACCCTT

CCACTGGCCGAGACAGCGCTCCCTGCCGCCCCCGCAAAGCCGCACGCGTGCATGTGGATG

CCGGCGCTGCTGAAGGTGCCGCTGGATGTCGCGGAAGGGATGATCAAAGTCACCAAGGAC

TTCATCGCCGCCCACCCGGAGGGCACGGTGACACTGGTGCTGCCTGACTGGTCGGCGGTG

GCCAGCGACGAAATCACCGGGGTGGAGAAAGACATCTCGGCCGCCCTCCAGGTGAACTGT

GCGCTTCTCAAGGCGTATGGACTGCCGAGCTCGGTGAAGATCGTGACGGAGAACGAGGTC

ATTCTCGGCAACTGCGACGACTTCTGGGTGAGTGTAATCGGCATTGCACGCAAGAACCTG

CTGAGCCACGTCGAGGAGCTGTACGGGGGAGAGGTACGCAACGCTGGTCAGGTCATTGCT

GCCCTCATGCGCGTTGCCACGGCTCTGATGCTATCCGTCTCGCACGTCATCTCAACCTCG

CTGGATGGACACATCAACGCCTTTGCCCGCGAGTACACGAAGGAGCGGATTGACTGCGTG

CAGACGCTGGAGGGACGCATCCCGGCGCTGCACCGCCCTGGCGCCGCCCCAGCGGTGCTC

GGTGCCGACGACGTCCTATACCTGGACGACAATGACATGGACATCCGCCGTAAAATTAAG

AAGGCGTACTCGGCGCCGAACGAGGAGGCCAATCCGGTCATATCGGTTGCCCAGCACCTT

CTCGCACAGCATGGAGCCCTCAGCATCGAGCGCGGGGAGGCCAACGGTGGCAACGTAAGC

TACAACACGCCAGAGGCCCTCGTGGCAGACTGCGGCAGCGGCGCTCTCCACCCTGCCGAC

CTCAAGGCGGCGGTGCTACAGCTGCTGCTGGACCGGTCGGCGCAGGCGCGCGCGCTGCTC

AACGGCGAGCTCAAGAAGAATATGACCGTTCTTCGCAACGCGGAGAAGAAGATGGCAAAG

AAGAGGTAA


 * *GC% Content for gene: ** 61.44%

ATGAACACAG ATGAGCGCTA CAAGCTCCTG CGTTCCGTTG GTGAGGAGTG CATCCAGGAG TCCGAGCTGC GCAATCTCAT TGAGAAGAAG CCTCTGATCC GCTGCTATGA TGGTTTCGAG CCCAGCGGCC GCATGCACAT CGCCCAGGGG ATCTTCAAGG CCGTGAACGT CAACAAATGC ACCGCGGCGG GTTGCGAATT CGTGTTCTGG GTCGCGGACT GGTTTGCCCT TATGAACGAT AAGGTGGGCG GCGAGCTGGA GAAGATTCGC ATCGTGGGGC GTTACCTTAT CGAGGTGTGG AAGGCGGCGG GCATGGACAT GGATAAGGTG CTGTTTCTGT GGTCCTCTGA GGAGATCACC TCGCACGCCG ACACCTACTG GCGTATGGTT CTCGATATCG GCCGCCAGAA CACAATCGCA CGGATCAAGA AGTGTTGCAC GATCATGGGC AAGACGGAAG GCACACTCAC TGCTGCCCAG GTGCTGTACC CGTTGATGCA GTGCTGCGAC ATCTTTTTCT TGAAGGCGGA TATCTGCCAG CTGGGCCTTG ACCAACGTAA GGTGAACATG CTCGCCCGCG AGTACTGTGA CCTCATTGGC CGCAAGCTTA AGCCCGTGAT TTTGTCCCAT CATATGCTCG CCGGGCTTAA GCAGGGCCAG GCGAAGATGT CGAAGAGCGA TCCCGACTCG GCGATTTTCA TGGAGGACAC GGAGGAGGAC GTGGCCCGGA AGATCCGCCA GGCCTACTGC CCCCGCGTGA AGCAGTCCGC GTCGGCCATC ACTGATGACG GCGCGCCGGT GGCGACAGAT GACCGCAACC CGGTGCTGGA CTATTTCCAG TGCGTGGTGT ACGCGCGCCC CGGTGCGGCG GCCACGATCG ACGGCACTAC GTACGCAACG TACGAGGACC TGGAGCAGGC GTTCGTTAGC GACGAGGTGA GCGAGGACGC GCTCAAGTCG TGCCTGATCG ATGAGGTCAA CGCACTGCTC GAGCCCGTCC GGCAGCACTT CGCCTCTAAC GAGGAAGCGC ATGAGCTGCT GGAGGCCGTG AAAAGCTACC GGAAGGACGG TGCGACACTC CCGCTGGCCG AGACGGCGCT CCCGGCGGCC CCCGCTAAGC CGCACGCGTG CATGTGGATG CCGGCCCTTC TGAAGGTGCC GCTGGATGTT GCAGAAGGTA TGATCAAGGT GACTAAGGAC TTCATCGCAG CGCACCCTGA AGGCACAGTG ACGCTGGTCC TCCCTGATTG GTCCGCCGTG GCGAGCGACG AGATCACCGG CGTCGAGAAG GATATCAGCG CGGCTCTGCA AGTTAATTGC GCTCTCCTCA AGGCGTACGG CCTGCCATCC AGCGTGAAGA TCGTGACAGA GAATGAGGTC ATCCTGGGCA ACTGCGACGA CTTCTGGGTC TCTGTCATCG GGATTGCACG CAAGAACCTG CTCAGCCATG TGGAGGAACT GTACGGCGGC GAGGTGCGCA ACGCCGGCCA GGTGATCGCG GCACTGATGC GCGTGGCCAC AGCCCTGATG CTGAGCGTCA GCCATGTTAT TTCTACCTCT CTGGACGGCC ACATTAATGC CTTCGCGCGC GAATACACGA AGGAGCGCAT CGACTGCGTG CAGACGCTGG AGGGTCGTAT CCCGGCGCTG CACCGGCCCG GCGCCGCGCC TGCCGTGCTC GGCGCGGACG ACGTGTTGTA CCTGGACGAC AACGACATGG ACATCCGTCG CAAGATCAAG AAGGCCTACA GCGCTCCGAA CGAGGAAGCC AACCCCGTCA TCTCCGTGGC CCAGCATCTC CTGGCGCAAC ATGGCGCGCT GTCCATTGAG CGCGGGGAGG CCAACGGCGG CAACGTGTCG TACAACACGC CGGAGGCCCT GGTGGCGGAC TGCGGCAGCG GTGCGCTCCA CCCCGCCGAT CTCAAGGCTG CGGTGCTCCA GCTTTTGCTG GATCGTTCTG CACAGGCTCG TGCGCTCCTC AACGGTGAGC TGAAGAAGAA TATGACGGTG CTTCGCAATG CCGAGAAGAA GATGGCGAAG AAGCGCTAA
 * *CDS Gene Sequence (codon optimized) - copy from ** **output** ** of ** **Primer Design** ** Protocol (paste as text only): **


 * *GC% Content for gene (codon optimized): ** 61.2 %

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


 * 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)

-- 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 'tail' primers (this is just 2 sequences): **