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.
Part of The Investigator's Annexe program.
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.
Part of The Investigator's Annexe program.
| Catalog Number | Product | DataSheet | Size | AVAILABILITY | Price | Qty |
|---|
| Product Type: | Protein |
| Name: | Histidyl-tRNA Synthetase Recombinant Protein w/6x His |
| Accession ID: | AAC75567, P12081 |
| 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 |
See Francklyn et al. Methods, 2008, for assay details
- Francklyn, C. and Schimmel, P., Enzymatic aminoacylation of an eight-base-pair microhelix with histidine. PNAS, 87, 8655-8659, (1990).
- Franckyln, C., Harris, D., and Moras, D., Crystallization of Histidyl-tRNA Synthase from Escherichia coli, Journal of Molecular Biology, 241, 275-277, (1994).
- 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).
- 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).
- 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).
- 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).
- 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).
- 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).
- 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).
- 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).
- 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).
- 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)
- 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).
- 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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