Changes in PLA2 activity after interacting with anti-inflammatory drugs and model membranes: evidence for the involvement of tryptophan residues

被引：14

作者：

Gaspar, Diana ^{[1
]}

Lucio, Marlene ^{[1
]}

Rocha, Sandra ^{[2
]}

Lima, J. L. F. Costa ^{[1
]}

Reis, Salette ^{[1
]}

机构：

[1] Univ Porto, Fac Farm, Dept Quim, REQUIMTE, P-4099030 Oporto, Portugal

[2] Univ Porto, Fac Engn, Dept Chem Engn, LEPAE, P-4200465 Oporto, Portugal

来源：

CHEMISTRY AND PHYSICS OF LIPIDS | 2011年 / 164卷 / 04期

关键词：

Liposomes; NSAIDs; PLA(2) inhibition; ADIFAB; Tryptophan fluorescence quenching and circular dichroism; SECRETORY PHOSPHOLIPASE A(2); INTERFACIAL BINDING SURFACE; PROTEIN SECONDARY STRUCTURE; FLUORESCENT-PROBE ADIFAB; FREE FATTY-ACIDS; CIRCULAR-DICHROISM; REGULATORY ROLE; EPC LIPOSOMES; ACTIVATION; INHIBITORS;

D O I：

10.1016/j.chemphyslip.2011.03.003

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Phospholipase A(2) (PLA(2)) lipolytic activity can be regarded as a limiting factor for the development of inflammatory processes by restricting the production of pro-inflammatory mediators, hence representing a valuable therapeutic target for drugs that are able to modulate the activity of this enzyme. In the current work, the hydrolysis of phospholipids by PLA(2) was monitored with acrylodan-labelled intestinal fatty acid binding protein (ADIFAB) and this fluorescence based technique was also used to access the enzymatic inhibitory effect of non-steroidal anti-inflammatory drugs (NSAIDs). The intrinsic fluorescence of PLA(2) tryptophan residues was further used to gain complementary information regarding the accessibility of these residues on the PLA(2) structure upon interaction with the NSAIDs tested; and to calculate the NSAIDs-PLA(2) binding constants. Finally, circular dichroism (CD) measurements were performed to evaluate changes in PLA(2) conformation resultant from the inhibitory effect of the drugs tested. Overall, results gathered in this study point to the conclusion that the studied NSAIDs inhibit PLA(2) activity due to a disturbance of the enzyme binding efficiency to membrane interface possibly by a shielding effect of the Trp residues required for the membrane interfacial binding step that precedes lipolysis process. (C) 2011 Elsevier Ireland Ltd. All rights reserved.

引用

页码：292 / 299

页数：8

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