Plasma Platelet-activating Factor Acetylhydrolase Is a Secreted Phospholipase A with a Catalytic Triad
Platelet-activating factor (PAF) is a potent pro-inflammatory autacoid with diverse physiological and pathological actions. These actions are modulated by PAF acetylhydrolase, which hydrolyzes the sn -2 ester bond to yield the biologically inactive lyso-PAF. In contrast to most secreted phospholipas...
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Veröffentlicht in: | The Journal of biological chemistry 1995-10, Vol.270 (43), p.25481 |
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Format: | Artikel |
Sprache: | eng |
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Zusammenfassung: | Platelet-activating factor (PAF) is a potent pro-inflammatory autacoid with diverse physiological and pathological actions.
These actions are modulated by PAF acetylhydrolase, which hydrolyzes the sn -2 ester bond to yield the biologically inactive lyso-PAF. In contrast to most secreted phospholipase A s, plasma PAF acetylhydrolase is calcium-independent and contains a G X S X G motif that is characteristic of the neutral lipases and serine esterases. In this study we tested whether the serine in
this motif is part of the active site of plasma PAF acetylhydrolase and, if so, what the other components of the active site
are. Using site-directed mutagenesis, we demonstrated that Ser-273 (of the G X S X G motif), Asp-296, and His-351 are essential for catalysis. These residues were conserved in PAF acetylhydrolase sequences
isolated from bovine, dog, mouse, and chicken. The linear orientation and spacing of these catalytic residues are consistent
with the α/β hydrolase conformation of other lipases and esterases. In support of this model, analysis of systematic truncations
of PAF acetylhydrolase revealed that deletions beyond 54 amino acids from the NH terminus and 21 from the COOH terminus resulted in a loss of enzyme activity. These observations demonstrate that although
plasma PAF acetylhydrolase is a phospholipase A it has structural properties characteristic of the neutral lipases and esterases. |
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ISSN: | 0021-9258 1083-351X |
DOI: | 10.1074/jbc.270.43.25481 |