Characterization, amino acid sequence and evolution of edema-inducing, basic phospholipase A2 from Trimeresurus flavoviridis venom.
Two phospholipases A2 (PLA2s) were purified from the venom of Trimeresurus flavoviridis (Crotalinae) inhabiting Tokunoshima island, Japan, and named PLA-A and PLA-B in the order of elution on a cation-exchange column. Lipolytic activities of PLA-A and PLA-B toward mixed micelles and liposomes were substantially lower than that of PLA2 (an [Asp49]PLA2) which had been isolated from the same venom. Both PLA-A and PLA-B consisted of 122 amino acids and contained aspartate at position 49 (the numbering according to the aligned sequences of PLA2s in Fig. 8), thus belonging to an [Asp49]PLA2 subgroup. PLA-A and PLA-B were identical in sequence with an exception at position 79. PLA-B contained Asn-Gly at positions 79 and 80 which are located in the beta-sheet region. On the other hand, PLA-A had beta-Asp-Gly and alpha-Asp-Gly in high and low proportion, respectively, at the corresponding positions which were produced from Asn-Gly through the base-catalyzed formation and hydrolysis of the succinimide type intermediate. Thus, PLA-A is derived from PLA-B. PLA-B is similar in sequence to PL-X, which had been purified from the venom of T. flavoviridis inhabiting Amami-Oshima island, Japan, and to PL-X', whose cDNA had been cloned from Tokunoshima T. flavoviridis venom gland, rather than PLA2. PLA-B showed strong edema-inducing activity, while PLA-A exhibited rather lower activity. The sequence around position 79 which constitutes a beta-turn segment seems to be crucial for edema-inducing activity. Phylogenetic tree of Tokunoshima T. flavoviridis venom PLA2 isozymes indicated that PLA-B and PL-X' diverged from PLA2 after branching of [Asp49]PLA2 forms and [Lys49]PLA2 forms.
Department of Chemistry, Faculty of Science, Kyushu University, Higashi-ku, 812-8581, Fukuoka, Japan.
Y
Y
Yamaguchi
Y
Y
Shimohigashi
T
T
Chijiwa
M
M
Nakai
T
T
Ogawa
S
S
Hattori
M
M
Ohno
D000595:Amino Acid Sequence; D000818:Animals; D005075:Biological Evolution; D002118:Calcium / Q000494:pharmacology; D003435:Crotalid Venoms / Q000201:enzymology*; D004487:Edema / Q000139:chemically induced* / Q000473:pathology; D004591:Electrophoresis, Polyacrylamide Gel; D008081:Liposomes; D008823:Micelles; D008969:Molecular Sequence Data; D010741:Phospholipases A / Q000032:analysis* / Q000633:toxicity*; D054467:Phospholipases A2; D010802:Phylogeny; D017841:Trimeresurus / Q000378:metabolism*
D003435:Crotalid Venoms; D008081:Liposomes; D008823:Micelles; D010741:Phospholipases A; D054467:Phospholipases A2; D002118:Calcium