Purification and characterization of a growth factor-like which increases capillary permeability from Vipera lebetina venom.
We have investigated the effect of Vipera lebetina venom on capillary permeability and isolated an increasing capillary permeability protein (ICPP) which is devoid of arginine ester hydrolase and phospholipase A2 activities. This protein was purified with a yield of about 0.2% by fast protein liquid chromatography (FPLC) using successively Superose 12, Mono Q, and Mono S columns and by high-pressure liquid chromatography (HPLC) on a C8 reverse-phase column. The purified protein migrated on SDS-PAGE as a band of about 27 kDa under nonreducing conditions and as a band of about 16 kDa under reducing conditions. Chromatography on a C8 column of reduced and alkylated protein yielded a single peak suggesting that this protein is homodimeric. This protein was refractory to Edman degradation chemistry. We used successfully a chemical unblocking involving the incubation of the protein with HCl in anhydrous methanol. The N-terminal amino acid sequence clearly shows considerable similarity to that of vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF).
Gasmi
A
A
Laboratoire Venins et Toxines, Institut Pasteur de Tunis, Tunis-Belvédère, Tunisia. ammar.gasmi@pasteur.rns.tn
Abidi
F
F
Srairi
N
N
Oijatayer
A
A
Karoui
H
H
Elayeb
M
M
D000595:Amino Acid Sequence; D000818:Animals; D002199:Capillary Permeability / Q000187:drug effects*; D019281:Dimerization; D016228:Endothelial Growth Factors / Q000235:genetics; D006133:Growth Substances / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology*; D008222:Lymphokines / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010982:Platelet-Derived Growth Factor / Q000235:genetics; D020836:Protein Structure, Quaternary; D017386:Sequence Homology, Amino Acid; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D014757:Viper Venoms / Q000737:chemistry* / Q000235:genetics / Q000633:toxicity; D017819:Viperidae
D016228:Endothelial Growth Factors; D006133:Growth Substances; D008222:Lymphokines; D010982:Platelet-Derived Growth Factor; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D014757:Viper Venoms