Glycan structures and intrageneric variations of venom acidic phospholipases A(2)  from Tropidolaemus pitvipers.
Most of the phospholipases A(2)  (PLA(2) ; EC3.1.1.4) variants isolated so far from snake venoms are nonglycosylated enzymes. In the present study, we purified an active glycosylated PLA(2)  and an inactive nonglycosylated Lys49-like PLA(2)  from two geographical venom samples of Tropidolaemus. The PLA(2)  variants from the two samples have rather different N-terminal sequences, implying that the samples were probably derived from two species (Tropidolaemus subannulatus and Tropidolaemus wagleri). The active PLA(2) s from Sulawesi and Sumatra venoms were designated as Tsu-E6 and Twa-E6, respectively, as a result of the presence of their conserved Glu6 residue. Tsu-E6 inhibited ADP-induced aggregation of mouse and human platelets. Twa-E6 stimulated the aggregation of mouse platelets but inhibited the aggregation of human platelets. Both PLA(2) s were found to be glycosylated at Asn14. Using MALDI-TOF analysis, the released glycans were shown to comprise complex type oligosaccharides without sialylation. This is the first glycan structure of the snake venom PLA(2)  to be solved. Furthermore, the enzymatic removal of glycans from both PLA(2) s did not significantly alter their effects on lipid hydrolysis and platelet aggregation. The thermostability of glycosylated Twa-E6 was also found to be as good as that of other homologous PLA(2) s. The presence of these oligosaccharides in PLA(2) s warrants further analyses, which may provide useful insights into the functional regulation of these biomolecules.
Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan. bc201@gate.sinica.edu.tw
Inn-Ho
IH
Tsai
Hui-Ching
HC
Chang
Jin-Mei
JM
Chen
An-Chun
AC
Cheng
Kay Hooi
KH
Khoo
D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D002152:Calorimetry, Differential Scanning; D003435:Crotalid Venoms / Q000737:chemistry* / Q000201:enzymology* / Q000235:genetics / Q000633:toxicity; D004795:Enzyme Stability; D014644:Genetic Variation; D006031:Glycosylation; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010974:Platelet Aggregation / Q000187:drug effects; D011134:Polysaccharides / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D017819:Viperidae / Q000235:genetics / Q000378:metabolism
D003435:Crotalid Venoms; D011134:Polysaccharides