Histidyl-tRNA synthetase (HARS) 6x-His

Full length E. coli Histidyl-tRNA synthetase (HARS) which has been purified using HisTrap affinity chromatography and contains a N-terminal 6x polyhistidine-tag.

Histidyl-tRNA synthetases attach amino acids to the 3' ends of their cognate transfer RNAs in a two-step reaction catalyzed by ATP. The accuracy of this reaction is crucial for the overall fidelity of protein synthesis. Defects in HARS are a cause of Usher syndrome type 3B (USH3B) [MIM:614504]. USH3B is a syndrome characterized by progressive vision and hearing loss during early childhood. Some patients have the so-called 'Charles Bonnet syndrome,' involving decreased visual acuity and vivid visual hallucinations. USH is a genetically heterogeneous condition characterized by the association of retinitis pigmentosa with sensorineural deafness. Age at onset and differences in auditory and vestibular function distinguish Usher syndrome type 1 (USH1), Usher syndrome type 2 (USH2) and Usher syndrome type 3 (USH3). USH3 is characterized by postlingual, progressive hearing loss, variable vestibular dysfunction, and onset of retinitis pigmentosa symptoms, including nyctalopia, constriction of the visual fields, and loss of central visual acuity, usually by the second decade of life.

From the laboratory of Christopher S. Francklyn, PhD, University of Vermont.

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Catalog Number Product Size AVAILABILITY Price Qty
Histidyl-tRNA synthetase (HARS) 6x-His, 50ug
50ug In stock
Regular Price:$256.00
Histidyl-tRNA synthetase (HARS) 6x-His, 100ug
100ug In stock
Regular Price:$410.00
Histidyl-tRNA synthetase (HARS) 6x-His, 250ug
250ug In stock
Regular Price:$769.00

Product Type: Protein
Name: Histidyl-tRNA Synthetase Recombinant Protein w/6x His
Accession ID: AAC75567, P60906
Source: E. coli
Molecular Weight: 47 kDa
Amino Acid Sequence: MGRSHHHHHHmakniqairgmndylpgetaiwqriegtlknvlgsygyseirlpiveqtplfkraigevtdvvekemytfedrngdsltlrpegtagcvragiehgllynqeqrlwyigpmfrherpqkgryrqfhqlgcevfglqgpdidaelimltarwwralgisehvtlelnsigslearanyrdalvafleqhkekldedckrrmytnplrvldsknpevqallndapalgdyldeesrehfaglckllesagiaytvnqrlvrgldyynrtvfewvtnslgsqgtvcaggrydglveqlggratpavgfamglerlvllvqavnpefkadpvvdiylvasgadtqsaamalaerlrdelpgvklmtnhgggnfkkqfaradkwgarvavvlgesevangtavvkdlrsgeqtavaqdsvaahlrtllg
Fusion Tag(s): N-terminal 6x His-tag
Purity: > 95% by HisTrap
Buffer: 50 mM HEPES pH 7.5; 100 mM KCl; 10 mM MgCl2, 5 mM ?-mercaptoethanol, 40% glycerol
Concentration: 100 uM
Storage: Store at -20C
Shipped: Dry ice

From the laboratory of Christopher S. Francklyn, PhD, University of Vermont.

See Francklyn et al. Methods, 2008, for assay details

  1. Francklyn, C. and Schimmel, P., Enzymatic aminoacylation of an eight-base-pair microhelix with histidine. PNAS, 87, 8655-8659, (1990).
  2. Franckyln, C., Harris, D., and Moras, D., Crystallization of Histidyl-tRNA Synthase from Escherichia coli, Journal of Molecular Biology, 241, 275-277, (1994).
  3. Yan, W. and Francklyn, C. Cytosine 73 is a discriminator nucleotide in vivo for histidyl-tRNA in Escherchia coli. Journal of Biological Chemistry, 269:13, 10022-10027 (1994).
  4. Bovee, M.L., Yan, W., Sproat, B.S.,  and Francklyn, C.S. tRNA discrimination at the binding step by a class II aminoacyl-tRNA synthetase. Biochemistry, 38: 41, 13725-13735 (1999).
  5. Arnez, J.G., Harris, D.C., Mitschler, A., Reese, B., Francklyn, C.S., and Moras, D.  Crystal structure of histidyl-tRNA synthetase from Escherichia coli complexed with histidyl-adenylate. EMBO Journal, 14:17, 4143-4155 (1995).
  6. Yan, W., Augustine, J., and Francklyn, C. A tRNA identity switch mediated by the binding interaction between a tRNA anticodon and the accessory domain of a class II aminoacyl-tRNA synthetase. Biochemistry, 35:21, 6559-6568 (1996).
  7. Arnez, J.G., Augustine, J.G., Moras, D., and Francklyn, C. The first step of aminoacylation at the atomic level in histidyl-tRNA synthase.  PNAS, 94, 7144-7149 (1997).
  8. Francklyn, C., Adams, J., and Augustine, J. Catalytic defects in mutants of class II histidyl-tRNA synthetase from Salmonella typhimurium previously linked to decreased control of histidine biosynthesis regulation.  Journal of Molecular Biology, 280, 847-858 (1998).
  9. Francklyn, C., Musier-Forsyth, K., and Martinis, S.A. Aminoacyl-tRNA synthetases in biology and disease: new evidence for structural and functional diversity in an ancient family of enzymes  RNA, 3, 945-960 (1997).
  10. Hawko, S.A., and Francklyn, C.S. Covariation of a specificity-determining structural motif in an aminoacyl-tRNA synthetase and a tRNA identity element. Biochemistry, 40:7, 1930-1936 (2001).
  11. Connoly, S.A., Rosen, A.E.,  Musier-Forsyth, K., and Francklyn, C.S. G-1:C73 recognition by an arginine cluster in the active site of Escherichia coli histidyl-tRNA synthetase.  Biochemistry, 43: 4, 962-969 (2004).
  12. Guth, E., Connoly, S.H., Bovee, M., and Francklyn, C.S. A substrate-assisted concerted mechanism for aminoacylation by a class II aminoacyl-tRNA synthetase. Biochemistry, 44:10, 3785-3794 (2005)
  13. Guth, E. and Francklyn, C.S. Kinetic discrimination of tRNA identity by the conserved motif 2 loop of a class II aminoacyl-tRNA synthetase. Molecular Cell, 25, 531-542 (2007).
  14. Francklyn, C.S., First, E.A., Perona, J.J., and Hou, Y. Methods for kinetic and thermodynamic analysis of aminoacyl-tRNA synthetases. Methods, 44:2, 100-118 (2008).

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