Synthesis and degradation of acyl peptide using enzyme from Pseudomonas aeruginosa.
The detailed properties of the enzyme from Pseudomonas aeruginosa, which catalyzes the N-acyl linkage between myristic acid and the N-terminal glycine residue of the octapeptide GNAAAARR-NH(2) (PKA) in aqueous solution without ATP and CoA, were studied. The substrate specificity for the acyl peptide in the synthetic reaction was examined, and it was found that at least eight amino acid residues are required for the reaction and that the N-terminal glycine residue is not absolutely essential for the reaction because the activity was detected using the octapeptide that has an N-terminal alanine. The activity was also strongly affected by the amino acid sequence because the activity was very weak in the reaction using GARASVLS-NH(2) (HIV-1p17(gag)). The substrate specificity for fatty acids was also examined. In the reactions using lauric acid and decanoic acid, only slight activities were detected; however, those activities were very small compared with the activity in the reaction using myristic acid. In addition, the degradation of myristoyl PKA by the enzyme was detected, although there are only a few reports on demyristoylation. The optimum pH and temperature of the degradation reaction were consistent with those of the synthetic reaction. The degradation reaction was inhibited by divalent cations.
Islam
Nazneen Naher
NN
Department of Applied Chemistry and Bioengineering, Graduate School of Engineering, Osaka City University, 3-3-138 Sugimoto, Sumiyoshi-ku, Osaka 558-8585, Japan.
Igarashi
Koichi
K
Tachibana
Taro
T
Ooshima
Hiroshi
H
Azuma
Masayuki
M
D000214:Acyl Coenzyme A / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000737:chemistry; D017868:Cyclic AMP-Dependent Protein Kinases / Q000737:chemistry; D005998:Glycine / Q000737:chemistry; D013058:Mass Spectrometry; D009227:Myristic Acids / Q000737:chemistry*; D009842:Oligopeptides / Q000737:chemistry*; D011550:Pseudomonas aeruginosa / Q000201:enzymology*; D013379:Substrate Specificity
D000214:Acyl Coenzyme A; D009227:Myristic Acids; D009842:Oligopeptides; D000255:Adenosine Triphosphate; D017868:Cyclic AMP-Dependent Protein Kinases; D005998:Glycine