Isolation and Functional Characterization of an Acidic Myotoxic Phospholipase A2 from Colombian Bothrops asper Venom

被引:13
作者
Posada Arias, Silvia [1 ,2 ]
Ray-Suarez, Paola [1 ]
Pereanez, Andres J. [1 ]
Acosta, Cristian [1 ]
Rojas, Mauricio [3 ]
dos Santos, Lucilene Delazari [4 ,5 ]
Ferreira, Rui Seabra, Jr. [4 ,5 ]
Nunez, Vitelbina [1 ,6 ]
机构
[1] Univ Antioquia, Programa Ofidismo & Escorpionismo, Medellin 050010, Colombia
[2] Corp Univ Lasallista, Sch Vet Med, Res Grp Vet Med GIVET, Caldas Antioquia 055440, Colombia
[3] Univ Antioquia, Grp Inmunol Celular & Inmunogenet GICIG, Medellin 050010, Colombia
[4] Univ Estadual Paulista, UNESP, Ctr Study Venoms & Venomous Anim CEVAP, BR-18610307 Botucatu, SP, Brazil
[5] Botucatu Med Sch FMB, Grad Program Trop Dis, BR-18610307 Botucatu, SP, Brazil
[6] Univ Antioquia, Escuela Microbiol, Medellin 050010, Colombia
关键词
acidic myotoxic phospholipase A(2); Bothrops asper; edema; myotoxicity; snake venom; SNAKE-VENOM; 3-DIMENSIONAL STRUCTURES; ANTIPLATELET; SEQUENCE; PURIFICATION; RECOGNITION; MECHANISM; JARARACA; ERRORS;
D O I
10.3390/toxins9110342
中图分类号
TS2 [食品工业];
学科分类号
0832 ;
摘要
Myotoxic phospholipases A(2) (PLA(2)) are responsible for many clinical manifestations in envenomation by Bothrops snakes. A new myotoxic acidic Asp49 PLA(2) (BaCol PLA(2)) was isolated from Colombian Bothrops asper venom using reverse-phase high performance liquid chromatography (RP-HPLC). BaCol PLA(2) had a molecular mass of 14,180.69 Da (by mass spectrometry) and an isoelectric point of 4.4. The complete amino acid sequence was obtained by cDNA cloning (GenBank accession No. MF319968) and revealed a mature product of 124 amino acids with Asp at position 49. BaCol PLA(2) showed structural homology with other acidic PLA(2) isolated from Bothrops venoms, including a non-myotoxic PLA(2) from Costa Rican B. asper. In vitro studies showed cell membrane damage without exposure of phosphatidylserine, an early apoptosis hallmark. BaCol PLA(2) had high indirect hemolytic activity and moderate anticoagulant action. In mice, BaCol PLA(2) caused marked edema and myotoxicity, the latter seen as an increase in plasma creatine kinase and histological damage to gastrocnemius muscle fibers that included vacuolization and hyalinization necrosis of the sarcoplasm.
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页数:15
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