Hemolytic, but not cell-invasive activity, of adenylate cyclase toxin is selectively affected by differential fatty-acylation in Escherichia coli.
Adenylate cyclase toxin from Bordetella pertussis requires posttranslational acylation of lysine 983 for the ability to deliver its catalytic domain to the target cell interior and produce cyclic adenosine monophosphate (cell-invasive activity) and to form transmembrane channels (hemolytic activity). When the toxin is expressed in Escherichia coli, it has reduced hemolytic activity, but comparable cell-invasive activity to that of adenylate cyclase toxin from B. pertussis. In contrast to the native protein from B. pertussis, which is exclusively palmitoylated, recombinant toxin from E. coli is acylated at lysine 983 with about 87% palmitoylated and the remainder myristoylated. Furthermore, the recombinant toxin contains an additional palmitoylation on approximately two-thirds of the lysines at position 860. These observations suggest that the site and nature of posttranslational fatty-acylation can be dictated by the bacterial host used for expression and can have a significant, but selective, effect on protein function.
Hackett
M
M
Department of Chemistry, University of Virginia, Charlottesville 22901, USA.
Walker
C B
CB
Guo
L
L
Gray
M C
MC
Van Cuyk
S
S
Ullmann
A
A
Shabanowitz
J
J
Hunt
D F
DF
Hewlett
E L
EL
Sebo
P
P
D000215:Acylation; D037361:Adenylate Cyclase Toxin; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001886:Bordetella pertussis / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D004912:Erythrocytes / Q000187:drug effects; D004926:Escherichia coli / Q000378:metabolism*; D006461:Hemolysis; D008239:Lysine; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D019308:Palmitic Acid; D010169:Palmitic Acids / Q000378:metabolism*; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism / Q000633:toxicity; D012756:Sheep; D010566:Virulence Factors, Bordetella / Q000096:biosynthesis / Q000378:metabolism* / Q000633:toxicity*
D037361:Adenylate Cyclase Toxin; D010169:Palmitic Acids; D010446:Peptide Fragments; D011994:Recombinant Proteins; D010566:Virulence Factors, Bordetella; D019308:Palmitic Acid; D008239:Lysine