title: N-myristylation of the catalytic subunit of cAMP-dependent protein kinase conveys structural stability.
abstract: Coexpression of the yeast N-myristyltransferase with the murine catalytic subunit of cAMP-dependent protein kinase in prokaryotic cells results in the N-myristylation of the recombinant catalytic subunit. The acylated recombinant catalytic subunit was purified following in vitro holoenzyme formation with a mutant form of the regulatory subunit and compared to the non-myristylated recombinant enzyme and to the mammalian porcine enzyme. All three enzymes are very similar in terms of their kinetic properties and their capacity to reassociate in vitro with the regulatory subunit to form holoenzyme. In contrast, the myristylated recombinant catalytic subunit is significantly more stable to thermal denaturation than the non-myristylated enzyme. Its thermal stability is now comparable to the mammalian enzyme. All three catalytic subunits are significantly more stable to thermal denaturation when they are part of the holoenzyme complex. Each shows an increase in T1/2 of 10 degrees C. This study demonstrates that one function for the myristic acid at the NH2 terminus of the catalytic subunit is to provide structural stability.
authors.affiliation: Department of Chemistry, University of California, San Diego, La Jolla 92093-0654.
authors.forename: W
authors.initials: W
authors.lastname: Yonemoto
authors.forename: M L
authors.initials: ML
authors.lastname: McGlone
authors.forename: S S
authors.initials: SS
authors.lastname: Taylor
mesh_terms: D000217:Acyltransferases / Q000378:metabolism*; D000818:Animals; D004926:Escherichia coli; D006358:Hot Temperature; D066298:In Vitro Techniques; D007700:Kinetics; D051379:Mice; D009226:Myristates / Q000378:metabolism*; D011489:Protein Denaturation; D011494:Protein Kinases / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology
chemical_list: D009226:Myristates; D011994:Recombinant Proteins; D000217:Acyltransferases; C050480:glycylpeptide N-tetradecanoyltransferase; D011494:Protein Kinases