Detergent binding to unmyristylated protein kinase A--structural implications for the role of myristate.
Myristylation often governs the targeting of protein kinases to the plasma membrane. It is now known that a key member of the src family of protein tyrosine kinases, pp60v-src, binds to the lipid bilayer of the plasma membrane via a myristylated amino terminal sequence. The mechanism of this interaction is not known; however, myristic acid (Myristic acid may also be referred to as Myristate) and residues 2 through 14 are also absolutely required (Resh and Ling, 1990). This review presents an analysis of crystal structures of detergent-modified recombinant and myristylated mammalian catalytic subunit of protein kinase A. Crystals of unmyristylated recombinant catalytic subunit of protein kinase A are grown in the presence of Mega 8, a glucamide-type of detergent, and only this detergent binds, which results in a resolution extension (Knighton et al., 1991a). Comparisons of these two structures reveal that the detergent association with the recombinant enzyme binds in exactly the same hydrophobic pocket of the protein occupied by myristic acid in the mammalian protein (Karlsson et al., 1993; Zheng et al., 1993a). Removal of the detergent through soaking results in the local unwinding of the first helix, helix A, and disorder of the canonical recognition sequence of the phosphorylation site, Ser 10 (Zheng et al., 1993b). These results suggest that anchoring the myristic acid inside the protein results in formation of a stable structural template, which includes the myristylated amino terminal sequence important for the recognition by protein kinases. This "inside out" motif might provide a structural paradigm for the recognition of myristylated proteins, including pp60v-src.
Sowadski
J M
JM
Department of Medicine, University of California at San Diego, La Jolla 92093-0654, USA.
Ellis
C A
CA
Madhusudan
D000818:Animals; D001665:Binding Sites; D002240:Carbohydrate Sequence; D002462:Cell Membrane / Q000201:enzymology; D018360:Crystallography, X-Ray; D017868:Cyclic AMP-Dependent Protein Kinases / Q000737:chemistry* / Q000378:metabolism*; D003902:Detergents / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015394:Molecular Structure; D019814:Myristic Acid; D009227:Myristic Acids / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011499:Protein Processing, Post-Translational
D003902:Detergents; D009227:Myristic Acids; D019814:Myristic Acid; D017868:Cyclic AMP-Dependent Protein Kinases