Characterization of three edema-inducing phospholipase A2 enzymes from habu (Trimeresurus flavoviridis) venom and their interaction with the alkaloid aristolochic acid

doi:10.1016/0041-0101(87)90286-8

Toxicon

Volume 25, Issue 5, 1987, Pages 501-515

https://doi.org/10.1016/0041-0101(87)90286-8 Get rights and content

Abstract

A basic phospholipase A₂ (PLA₂) enzyme, TFV PL-X (pI 9.2) and two acidic PLA₂ enzymes, TFV PL-Ia (pI 4.9) and TFV PL-Ib (pI 4.5) were purified from Trimeresurus flavoridis venom on CM-Sephadex C-25 and QAE-Sephadex A-25 columns, respectively. The basic enzyme exists as a monomer, whereas the acidic enzymes are dimers. These enzymes differ in properties such as molecular weight, K_m, optimum pH and temperature and pharmacological properties. The basic enzyme hydrolysed purified phospholipids in the order of PC > PE > PS > PI = 0, while for TFV PL-Ia and TFV PL-Ib the order was PC > PE > PS = PI = 0. TFV PL-X was comparatively more toxic, with an ld₅₀ value of 4.2 μg/g (i.p.), while the acidic PLA₂ enzymes had ld₅₀ values above 8 μg/g (i.p.). All three enzymes induced edema when injected into the mouse foot pad.

Aristolochic acid, an alkaloid (8-methoxy-6-nitrophenanthro(3,4-d)-1,3-dioxole-5-carboxylic acid) from the medicinal plant Aristolochia radix, interacts with these PLA₂ enzymes. It is a competitive inhibitor with varying affinity when PC is used as substrate. Aristolochic acid inhibits direct and indirect hemolytic activity, as well as edema-inducing activity, of TFV PL-X, but fails to neutralize the lethal potentcy of the enzyme. On the other hand, it inhibits direct and indirect lytic activity of TFV PL-Ia and TFV PL-Ib only at 10-fold higher concentrations and it enhances the edema-inducing activity of these enzymes. Such effects of aristolochic acid indicates that (1) different mechanisms may be involved in the edema-inducing activity of PLA₂ enzymes and (2) catalytic and pharmacological sites are separate on the PLA₂ molecule.

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^☆: Part of the work was accepted for presentation at the 13th International Congress of Biochemistry held 25–30 August 1985 in Amsterdam, The Netherlands.

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Characterization of three edema-inducing phospholipase A2 enzymes from habu (Trimeresurus flavoviridis) venom and their interaction with the alkaloid aristolochic acid☆

Abstract

J. biol. Chem.

Biochim. biophys. Acta

Biochem. Pharmac.

Biochem. Pharmac.

Toxicon

Toxicon

Toxicon

Toxicon

Toxicon

Biochem. biophys. Res. Commun.

Biochim. biophys. Acta

Toxicon

Toxicon

Insect Biochem.

J. biol. Chem.

Analyt. Biochem.

Estimation of the molecular weights of proteins by Sephadex gel-filtration

Biochem. J.

A comparative study of the interaction of chlorpromazine, trifluoperazine, and promethazine with mouse brain tubulin

Molec. Pharmac.

Characterization of three edema-inducing phospholipase A₂ enzymes from habu (Trimeresurus flavoviridis) venom and their interaction with the alkaloid aristolochic acid☆