Isolation of a polypeptide from Phoneutria nigriventer spider venom responsible for the increased vascular permeability in rabbit skin.

Fractionation of Phoneutria nigriventer venom by Sephadex G-10 followed by ion-exchange chromatography yields a fraction (fraction XIII) which increases microvascular permeability in rabbit skin in vivo by activating the tissue kallikrein-kinin system. One polypeptide (PNV3) with the ability to increase microvascular permeability in the rabbit skin in vivo was isolated from fraction XIII and biochemically characterized. PNV3 has 132 amino acid residues with a calculated mol. wt of 14,475. This polypeptide showed the following N-terminal sequence: AVFAIQDQPC. Amino acid analysis indicated the presence of six disulfide bridges and a high content of Glx (20%). Pairwise comparison of PNV3 amino acid sequence with 27 other spider venom polypeptides and proteins indicated that PNV3 presents high similarity (60-70%) with other toxins (Tx2.1, Tx2.5 and Tx2.6) isolated from P. nigriventer venom.

Metabolic behavior in rats of a nonprotein microemulsion resembling low-density lipoprotein.

A protein-free microemulsion (LDE) with a lipid composition resembling that of low-density lipoprotein (LDL) was used in metabolic studies in rats to compare LDE with the native lipoprotein. LDE labeled with radioactive lipids was injected into the bloodstream of male Wistar rats, and plasma kinetics of the labeled lipids were followed on plasma samples collected at regular intervals for 12 h after injection. The 24-h LDE uptake by different tissues was also measured in tissue samples excised after the animals had been sacrificed. We found that LDE plasma kinetics were similar to those described for native LDL [fractional clearance rate (FCR) of cholesteryl ester, 0.42 +/- 0.11 h-1]. The major site for LDE uptake was the liver, and the tissue distribution of the LDE injected radioactivity was as one would expect for LDL. To test whether LDE was taken up by the specific LDL receptors, the LDE emulsion was injected into rats treated with 17 alpha-ethinylestradiol, which is known to increase the activity of these receptors; as expected, removal of LDE from the bloodstream increased (FCR = 0.90 +/- 0.35 h-1). On the other hand, saturation of the receptors that remove remnants by prior infusion of massive amounts of lymph chylomicrons did not change LDE plasma kinetics. These results indicate that LDE is cleared from plasma by B,E receptors and not by the E receptors that remove remnants. Incorporation of free cholesterol into LDE increased LDE plasma clearance. Incubation studies also showed that LDE incorporates a variety of apolipoproteins, including apo E, a ligand for recognition of lipoproteins by specific receptors.(ABSTRACT TRUNCATED AT 250 WORDS)