title: Expression, purification, and buffer solubility optimization of the putative human peptidyl-tRNA hydrolase PTRHD1.
abstract: Performing the essential function of recycling peptidyl-tRNAs, peptidyl-tRNA hydrolases are ubiquitous in all domains of life. The multicomponent eukaryotic Pth system differs greatly from the bacterial system composed predominantly of a single Pth1 enzyme. While bacterial Pth1s are structurally well characterized and promising new targets for antibiotic development, eukaryotic Pths are largely understudied. From amino acid sequence alignment and secondary structure predictions, the human gene product PTRHD1 was classified as a eukaryotic Pth. Herein, we report cloning, recombinant bacterial expression, and weak binding to peptidyl-tRNA for PTRHD1. Additionally, we report binding to tRNA but absence of peptidyl-tRNA hydrolase activity. Thus, PTRHD1 is not a Pth and the functional consequence of nucleotide binding remains undefined.
authors.affiliation: Department of Chemistry, The University of Alabama in Huntsville, 301 Sparkman Drive, Huntsville, AL 35899, USA.
authors.forename: Geordan L
authors.initials: GL
authors.lastname: Burks
authors.affiliation: Department of Chemistry, The University of Alabama in Huntsville, 301 Sparkman Drive, Huntsville, AL 35899, USA.
authors.forename: Hana
authors.initials: H
authors.lastname: McFeeters
authors.affiliation: Department of Chemistry, The University of Alabama in Huntsville, 301 Sparkman Drive, Huntsville, AL 35899, USA. Electronic address: robert.mcfeeters@uah.edu.
authors.forename: Robert L
authors.initials: RL
authors.lastname: McFeeters
mesh_terms: D002265:Carboxylic Ester Hydrolases / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D011994:Recombinant Proteins; D012995:Solubility
chemical_list: D011994:Recombinant Proteins; D002265:Carboxylic Ester Hydrolases; C019674:aminoacyl-tRNA hydrolase
keywords: C2orf79; PTRHD1; Peptidyl-tRNA hydrolase domain containing 1; Purification; Recombinant expression; Solubility optimization