Target+-+Tyrosyl-tRNA+Synthetase+-+L.+Major


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


 * *NCBI Gene # or RefSeq#: **27213


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


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


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


 * *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. It is transmitted by the female sandfly and can cause uncomfortable skin lesions.


 * 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
 * <span style="font-family: 'Calibri','sans-serif';">Essentiality of this protein: **


 * <span style="font-family: 'Calibri','sans-serif';">Complex of proteins?: **<span style="font-family: 'Calibri','sans-serif';">yes

Synthetic analogues of SB-219383. Novel C-glycosyl peptides as inhibitors of tyrosyl tRNA synthetase. (2001).
 * <span style="font-family: 'Calibri','sans-serif';">Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): ****<span style="font-family: 'Calibri','sans-serif';">Homo sapiens (1) **


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


 * <span style="font-family: 'Calibri','sans-serif';">*EC#: **<span style="font-family: 'Calibri','sans-serif';">6.1.1.1


 * <span style="font-family: 'Calibri','sans-serif';">Link to BRENDA EC# page: **<span style="font-family: 'Calibri','sans-serif';">http://brenda-enzymes.org/php/result_flat.php4?ecno=6.1.1.1

<span style="font-family: 'Calibri','sans-serif';">

**Enzyme Assay** **<span style="font-family: 'Calibri','sans-serif';"> 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


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

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


 * <span style="font-family: 'Calibri','sans-serif';">Current Inhibitors: **


 * <span style="font-family: 'Calibri','sans-serif';">Expression Information (has it been expressed in bacterial cells): **<span style="font-family: 'Calibri','sans-serif';"> Yes, expressed in E. coli.

**Purification Method** **<span style="font-family: 'Calibri','sans-serif';">: **<span style="font-family: 'Calibri','sans-serif';">Ni-NTA affinity chromatography <span style="font-family: 'Calibri','sans-serif';">

MNTDERYKLLRSVGEECIQESELRNLIEKKPLIRCYDGFEPSGRMHIAQGIFKAVNVNKC
 * <span style="font-family: 'Calibri','sans-serif';">Image of protein (PyMol with features delineated and shown separately): **
 * <span style="font-family: 'Calibri','sans-serif';">*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): **

TAAGCEFVFWVADWFALMNDKVGGELEKIRIVGRYLIEVWKAAGMDMDKVLFLWSSEEIT

SHADTYWRMVLDIGRQNTIARIKKCCTIMGKTEGTLTAAQVLYPLMQCCDIFFLKADICQ

LGLDQRKVNMLAREYCDLIGRKLKPVILSHHMLAGLKQGQAKMSKSDPDSAIFMEDTEED

VARKIRQAYCPRVKQSASAITDDGAPVATDDRNPVLDYFQCVVYARPGAAATIDGTTYAT

YEDLEQAFVSDEVSEDALKSCLIDEVNALLEPVRQHFASNEEAHELLEAVKSYRKDGATL

PLAETALPAAPAKPHACMWMPALLKVPLDVAEGMIKVTKDFIAAHPEGTVTLVLPDWSAV

ASDEITGVEKDISAALQVNCALLKAYGLPSSVKIVTENEVILGNCDDFWVSVIGIARKNL

LSHVEELYGGEVRNAGQVIAALMRVATALMLSVSHVISTSLDGHINAFAREYTKERIDCV

QTLEGRIPALHRPGAAPAVLGADDVLYLDDNDMDIRRKIKKAYSAPNEEANPVISVAQHL

LAQHGALSIERGEANGGNVSYNTPEALVADCGSGALHPADLKAAVLQLLLDRSAQARALL

NGELKKNMTVLRNAEKKMAKKR


 * <span style="font-family: 'Calibri','sans-serif';">*length of your protein in Amino Acids: **<span style="font-family: 'Calibri','sans-serif';">682aa

**Molecular Weight** **<span style="font-family: 'Calibri','sans-serif';">: **<span style="font-family: 'Calibri','sans-serif';">74.949 kDa


 * <span style="font-family: 'Calibri','sans-serif';">Molar ** **Extinction coefficient** **<span style="font-family: 'Calibri','sans-serif';"> of your protein at 280 nm wavelength: **<span style="font-family: 'Calibri','sans-serif';">71945


 * <span style="color: #000000; font-family: 'Calibri','sans-serif';">TMpred graph Image **


 * <span style="font-family: 'Calibri','sans-serif';">*CDS ** **Gene Sequence** **<span style="font-family: 'Calibri','sans-serif';"> (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


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


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


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

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


 * <span style="font-family: 'Calibri','sans-serif';">Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): **


 * <span style="font-family: 'Calibri','sans-serif';">( <span style="color: #008000; font-family: 'Calibri','sans-serif';">link to DNA Works output ** ** text file ** **<span style="color: #008000; font-family: 'Calibri','sans-serif';"> - **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.


 * <span style="font-family: 'Calibri','sans-serif';">Primer design results for 'tail' primers (this is just 2 sequences): **