RESUMO
Methods for cell-selective analysis of proteome dynamics will facilitate studies of biological processes in multicellular organisms. Here we describe a mutant murine methionyl-tRNA synthetase (designated L274GMmMetRS) that charges the noncanonical amino acid azidonorleucine (Anl) to elongator tRNA(Met) in hamster (CHO), monkey (COS7), and human (HeLa) cell lines. Proteins made in cells that express the synthetase can be labeled with Anl, tagged with dyes or affinity reagents, and enriched on affinity resin to facilitate identification by mass spectrometry. The method does not require expression of orthogonal tRNAs or depletion of canonical amino acids. Successful labeling of proteins with Anl in several mammalian cell lines demonstrates the utility of L274GMmMetRS as a tool for cell-selective analysis of mammalian protein synthesis.
Assuntos
Aminoacil-tRNA Sintetases/metabolismo , Biossíntese de Proteínas , Aminoácidos/análise , Aminoácidos/química , Aminoacil-tRNA Sintetases/genética , Animais , Cricetinae , Escherichia coli/metabolismo , Haplorrinos , Células HeLa , Humanos , Mamíferos , Metionina tRNA Ligase/metabolismo , Camundongos , Proteoma/metabolismo , RNA de Transferência de Metionina/metabolismoRESUMO
Firearm injuries are a leading cause of pediatric mortality in the United States. The frequency of pediatric extremity firearm injuries and the high repeat incidence in high-risk patients make it important for emergency clinicians to understand how to manage these injuries. This issue focuses on the acute management of firearm injuries to the extremities of pediatric patients, drawing from the pediatric literature or extrapolated from adult literature where pediatric evidence is scarce. Current trends, novel management, and controversies are also discussed.