The first representative of glycosylated three-fingered toxins. Cytotoxin from the Naja kaouthia cobra venom.

Publisher: Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies Country of Publication: England NLM ID: 0107600 Publication Model: Print Cited Medium: Print ISSN: 0014-2956 (Print) Linking ISSN: 00142956 NLM ISO Abbreviation: Eur J Biochem Subsets: MEDLINE

Publication: -2004: Oxford, UK : Blackwell Science Ltd. on behalf of the Federation of European Biochemical Societies
Original Publication: Berlin, New York, Springer.

Cytotoxins/*chemistry ; Elapid Venoms/*chemistry ; Glycoproteins/*chemistry; Amino Acid Sequence ; Animals ; Binding Sites ; Circular Dichroism ; Consensus Sequence ; Cytotoxins/isolation & purification ; Cytotoxins/toxicity ; Elapidae/metabolism ; Glucans/chemistry ; Glycoproteins/isolation & purification ; Glycoproteins/toxicity ; Glycosylation ; Gryllidae/drug effects ; HL-60 Cells ; Humans ; Models, Molecular ; Molecular Sequence Data ; Peptide Fragments/chemistry ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Trypsin/metabolism

There are different glycosylated proteins in snake venoms, but no glycosylated representatives of a large family of three-fingered toxins have previously been detected. A new glycoprotein was isolated from the venom of the Thai cobra Naja kaouthia. MALDI MS of the glycoprotein contained an array of peaks in the range from approximately 8900 to approximately 9400 Da indicating its microheterogeneity. Carbohydrate analysis showed the presence of mannose, galactose, N-acetylglucosamine, fucose and neuraminic acid. The N-terminal sequence of the glycoprotein was identical to that of cytotoxin 3 (CX3) from N. kaouthia, and CD spectra of the glycoprotein and CX3 were almost the same. Cleavage of a glycan moiety by N-glycosidase F gave a protein of molecular mass practically coinciding with that of CX3. MALDI MS of the tryptic digest of reduced glycoprotein S-pyridylethylated at cysteine residues, contained peaks corresponding to all tryptic fragments of CX3, with the exception of fragment 24-30. The peak corresponding to this peptide appeared in the mass-spectrum of similarly treated deglycosylated glycoprotein. These data show that the potential N-glycosylation site at Asn29 in CX3 is utilized for glycan attachment and that the glycoprotein is glycosylated CX3. In vivo toxicity of the glycoprotein to the cricket Gryllus assimilis was twofold lower than that of CX3. The cytotoxic activity of the glycoprotein towards HL60 cells was about two orders of magnitude lower than that of CX3, but could be made equal to the CX3 cytotoxicity by deglycosylation. Thus for the first time we have isolated a glycosylated three-fingered snake venom toxin wherein glycosylation appears to modulate its biological activity.

0 (Cytotoxins)
0 (Elapid Venoms)
0 (Glucans)
0 (Glycoproteins)
0 (Peptide Fragments)
EC 3.4.21.4 (Trypsin)

Date Created: 20040507 Date Completed: 20040713 Latest Revision: 20171116

20231215

10.1111/j.1432-1033.2004.04115.x

15128311