title: Expression, purification, and characterization of rhTyrRS.
abstract: BACKGROUND
  Aminoacyl-tRNA synthetases (AARSs) catalyze the first step of protein synthesis. Emerging evidence indicates that AARSs may have additional functions, playing a role in signal transduction pathways regulating thrombopoiesis and inflammation. Recombinant human tyrosyl-tRNA synthetase (rhTyrRS) is engineered with a single amino acid substitution that unmasks its cytokine activity. An industrial production method that provides high yield as well as high purity, quality, and potency of this protein is required for preclinical research.
  
  
  RESULTS
  We expressed codon-optimized rhTyrRS in Escherichia coli under fermentation conditions. Soluble protein was purified by a three-step purification method using cation exchange chromatography, gel filtration chromatography, and anion exchange chromatography. We also established a method to test the biological activity of rhTyrRS by measuring aminoacylation and IL-8 release in rhTyrRS-treated HL-60 cells.
  
  
  CONCLUSIONS
  The characterization of purified rhTyrRS indicated that this protein can be used in pharmacodynamic and pharmacokinetic studies.
authors.forename: Yongjiang
authors.initials: Y
authors.lastname: Lang
authors.forename: Yanling
authors.initials: Y
authors.lastname: Zhang
authors.forename: Ling
authors.initials: L
authors.lastname: Zhan
authors.forename: Zhe
authors.initials: Z
authors.lastname: Feng
authors.forename: Xiushi
authors.initials: X
authors.lastname: Zhou
authors.affiliation: The Key Laboratory of Molecular Medicine, Ministry of Education, Fudan University, Shanghai, P,R, China. minyu@shmu.edu.cn.
authors.forename: Min
authors.initials: M
authors.lastname: Yu
authors.forename: Wei
authors.initials: W
authors.lastname: Mo
mesh_terms: D000215:Acylation; D002851:Chromatography, High Pressure Liquid; D056148:Chromatography, Reverse-Phase; D004926:Escherichia coli / Q000378:metabolism; D015870:Gene Expression; D018922:HL-60 Cells; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism; D013058:Mass Spectrometry; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D014448:Tyrosine-tRNA Ligase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D016209:Interleukin-8; D011994:Recombinant Proteins; D014448:Tyrosine-tRNA Ligase