title: Myristoylome profiling reveals a concerted mechanism of ARF GTPase deacylation by the bacterial protease IpaJ.
abstract: N-myristoylation is an essential fatty acid modification that governs the localization and activity of cell signaling enzymes, architectural proteins, and immune regulatory factors. Despite its importance in health and disease, there are currently no methods for reversing protein myristoylation in vivo. Recently, the Shigella flexneri protease IpaJ was found to cleave myristoylated glycine of eukaryotic proteins, yet the discriminatory mechanisms of substrate selection required for targeted demyristoylation have not yet been evaluated. Here, we performed global myristoylome profiling of cells treated with IpaJ under distinct physiological conditions. The protease is highly promiscuous among diverse N-myristoylated proteins in vitro but is remarkably specific to Golgi-associated ARF/ARL family GTPases during Shigella infection. Reconstitution studies revealed a mechanistic framework for substrate discrimination based on IpaJ's function as a GTPase "effector" of bacterial origin. We now propose a concerted model for IpaJ function that highlights its potential for programmable demyristoylation in vivo.
authors.affiliation: Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, TX 75390-8816, USA.
authors.forename: Nikolay
authors.initials: N
authors.lastname: Burnaevskiy
authors.affiliation: Laboratory of Chemical Biology and Microbial Pathogenesis, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Tao
authors.initials: T
authors.lastname: Peng
authors.affiliation: Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, TX 75390-8816, USA.
authors.forename: L Evan
authors.initials: LE
authors.lastname: Reddick
authors.affiliation: Laboratory of Chemical Biology and Microbial Pathogenesis, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Howard C
authors.initials: HC
authors.lastname: Hang
authors.affiliation: Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, TX 75390-8816, USA. Electronic address: neal.alto@utsouthwestern.edu.
authors.forename: Neal M
authors.initials: NM
authors.lastname: Alto
mesh_terms: D020823:ADP-Ribosylation Factor 1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000089982:ADP-Ribosylation Factor 6; D020727:ADP-Ribosylation Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000942:Antigens, Bacterial / Q000235:genetics / Q000378:metabolism*; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019814:Myristic Acid / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012763:Shigella flexneri / Q000737:chemistry* / Q000201:enzymology; D015398:Signal Transduction
chemical_list: D000089982:ADP-Ribosylation Factor 6; D000942:Antigens, Bacterial; C110952:IpaJ protein, Shigella flexneri; D011994:Recombinant Proteins; D019814:Myristic Acid; D020823:ADP-Ribosylation Factor 1; D020727:ADP-Ribosylation Factors