title: A prokaryote-based cell-free translation system that efficiently synthesizes glycoproteins.
abstract: Asparagine-linked (N-linked) protein glycosylation has been observed in all domains of life, including most recently in bacteria and is now widely considered a universal post-translational modification. However, cell-based production of homogeneous glycoproteins for laboratory and preparative purposes remains a significant challenge due in part to the complexity of this process in vivo. To address this issue, an easily available and highly controllable Escherichia coli-based cell-free system for the production of N-linked glycoproteins was developed. The method was created by coupling existing in vitro translation systems with an N-linked glycosylation pathway reconstituted from defined components. The translation/glycosylation system yielded efficiently glycosylated target proteins at a rate of hundreds of micrograms/milliliters in half a day. This is the first time a prokaryote-based cell-free protein synthesis system has generated N-linked glycoproteins.
authors: Comparative Biomedical Sciences, Cornell University, Ithaca, NY 14853, USA.
authors: Cassandra
authors: C
authors: Guarino
authors: Matthew P
authors: MP
authors: DeLisa
mesh_terms: D002474:Cell-Free System; D004926:Escherichia coli / Q000378:metabolism; D006023:Glycoproteins / Q000096:biosynthesis*; D006031:Glycosylation; D011387:Prokaryotic Cells / Q000378:metabolism*; D014176:Protein Biosynthesis
chemical_list: D006023:Glycoproteins