Characterization of an apoptosis-inducing factor in Habu snake venom as a glycyrrhizin (GL)-binding protein potently inhibited by GL in vitro.
By means of successive heparin-affinity and glycyrrhizin (GL)-affinity column chromatographies (HPLC), a 55 kDa GL-binding protein (gp55) was purified to apparent homogeneity from the Superdex P-I fraction of Habu snake venom. This gp55 was identified as an apoxin I-like protein, because (i) its 20 N-terminal amino acid residues (AHDRNPLEEYFRETDYEEFL) are 95% identical with the corresponding sequence of apoxin I (apoptosis-inducing factor, approx. 55 kDa) in the venom of the western diamondback rattlesnake; and (ii) L-amino acid oxidase (LAO) activity of gp55 is detected when incubated with L-leucine, but not with D-leucine. GL inhibited the LAO activity of gp55 in a dose-dependent manner, but had no effect on the activity of a 65 kDa LAO also purified from Habu snake venom. In addition, GL reduced the ability of gp55 to induce the hemolysis of sheep red blood cells. These results suggest that GL is a potent inhibitor of apoxin I-like proteins in harmful snake venoms.
Abe
Y
Y
Laboratory of Genetical Biochemistry, Kitasato University School of Allied Health Sciences, Sagamihara, Japan.
Shimoyama
Y
Y
Munakata
H
H
Ito
J
J
Nagata
N
N
Ohtsuki
K
K
D000594:Amino Acid Oxidoreductases / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000494:pharmacology*; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D003435:Crotalid Venoms / Q000737:chemistry*; D019695:Glycyrrhizic Acid / Q000378:metabolism* / Q000494:pharmacology; D006461:Hemolysis / Q000187:drug effects; D006493:Heparin; D050830:L-Amino Acid Oxidase; D008969:Molecular Sequence Data; D011485:Protein Binding; D012756:Sheep; D017841:Trimeresurus; D014757:Viper Venoms
D002352:Carrier Proteins; D003435:Crotalid Venoms; D014757:Viper Venoms; C490099:glycyrrhizin-binding protein, Trimeresurus flavoviridis; C086010:glycyrrhizin-binding proteins; D019695:Glycyrrhizic Acid; D006493:Heparin; D000594:Amino Acid Oxidoreductases; D050830:L-Amino Acid Oxidase