Inhibition of secretory phospholipase A(2) enzyme by bilirubin: a new role as endogenous anti-inflammatory molecule.
Bilirubin is a powerful antioxidant that suppresses the inflammatory process. However its interaction with proinflammatory PLA(2) enzyme is not known. Inhibition of several secretory phospholipase A(2) (sPLA(2)) enzyme activities by bilirubin was studied using (14)C-oleate labeled Escherichia coli as substrate. Bilirubin inhibits purified sPLA(2) enzyme from Vipera russellii and Naja naja venom and partially purified sPLA(2) enzymes from human ascitic fluid, pleural fluid and normal serum in a dose dependent manner. IC(50) values calculated for these enzymes ranges from 1.75 to 10.5 microM. Inflammatory human sPLA(2) enzymes are more sensitive to inhibition by bilirubin than snake venom sPLA(2)s. Inhibition of sPLA(2) activity by bilirubin is independent of calcium concentration. Increasing substrate concentration (upto 180 nmol) did not relieve the inhibition of sPLA(2) by bilirubin and it is irreversible. Bilirubin quenched the relative fluorescence intensity of sPLA(2) in a dose dependent manner in the same concentration range at which in vitro sPLA(2) inhibition was observed. In the presence of bilirubin, apparent shift in the far UV-CD spectra of sPLA(2) was observed, indicating a direct interaction with the enzyme. Inhibition of sPLA(2) induced mouse paw edema by bilirubin confirms its sPLA(2) inhibitory activity in vivo also. These findings indicate that inhibition of sPLA(2) by bilirubin is mediated by direct interaction with the enzyme and bilirubin may act as an endogenous regulator of sPLA(2) enzyme activity.
Jameel
Noor Mohamed
NM
Department of Studies in Biochemistry, University of Mysore, Manasagangothri, Mysore 570 006, India. jameelmd@yahoo.com
Frey
Brigitte M
BM
Frey
Felix J
FJ
Gowda
T Veerabasappa
TV
Vishwanath
Bannikuppe S
BS
D000818:Animals; D000893:Anti-Inflammatory Agents / Q000378:metabolism* / Q000494:pharmacology; D001202:Ascitic Fluid / Q000201:enzymology; D001663:Bilirubin / Q000378:metabolism* / Q000494:pharmacology*; D002118:Calcium / Q000494:pharmacology; D002417:Cattle; D004305:Dose-Response Relationship, Drug; D004487:Edema / Q000139:chemically induced; D006801:Humans; D020128:Inhibitory Concentration 50; D010741:Phospholipases A / Q000037:antagonists & inhibitors* / Q000378:metabolism; D035422:Pleural Cavity / Q000201:enzymology; D012910:Snake Venoms / Q000201:enzymology; D012911:Snakes; D013050:Spectrometry, Fluorescence
D000893:Anti-Inflammatory Agents; D012910:Snake Venoms; D010741:Phospholipases A; D001663:Bilirubin; D002118:Calcium