title: Coexpression of proteins with methionine aminopeptidase and/or N-myristoyltransferase in Escherichia coli to increase acylation and homogeneity of protein preparations.
abstract: New plasmid constructs described in this article allow the coexpression in bacteria of any protein with several different NMT proteins, including the recently cloned full-length human NMT1 and 2, and with increased expression of bacterial Met-AP. Through the use of these plasmids in different combinations it should be possible to improve the homogeneity of a large number of recombinant protein preparations by the complete removal of the initiating methionine and increased extent of N-myristoylation. The new reagents described in this article are available upon request.
authors.affiliation: GlaxoSmithKline, King of Prussia, Pennsylvania 19406, USA.
authors.forename: Hillary A
authors.initials: HA
authors.lastname: Van Valkenburgh
authors.forename: Richard A
authors.initials: RA
authors.lastname: Kahn
mesh_terms: D000215:Acylation; D000217:Acyltransferases / Q000235:genetics / Q000378:metabolism*; D000626:Aminopeptidases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D001483:Base Sequence; D017931:DNA Primers; D004926:Escherichia coli / Q000235:genetics*; D006801:Humans; D008322:Mammals; D063208:Methionyl Aminopeptidases; D019814:Myristic Acid / Q000378:metabolism; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection / Q000379:methods
chemical_list: D001426:Bacterial Proteins; D017931:DNA Primers; D011994:Recombinant Proteins; D019814:Myristic Acid; D000217:Acyltransferases; C050480:glycylpeptide N-tetradecanoyltransferase; D000626:Aminopeptidases; D063208:Methionyl Aminopeptidases