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Sea nettle (Chrysaora quinquecirrha) lethal factor: purification by recycling on m-aminophenyl boronic acid acrylic beads



Sea nettle (Chrysaora quinquecirrha) lethal factor: purification by recycling on m-aminophenyl boronic acid acrylic beads



Toxicon 32(4): 467-478



Experiments were performed to define the best isolation method for isolating Chrysaora fishing tentacle nematocyst organelles in order to minimize non-nematocyst contaminating proteins and proteases and stabilize crude nematocyst venom lethal activity. Techniques employed to disrupt the tentacles included autolysis, homogenization, or digestion using either trypsin or collagenase. Sephacryl-200 gel-filtration chromatography separated two lethal fractions. An immobilized serine protease inhibitor column, m-aminophenyl boronic acid acrylic beads, which reversibly bound one of the two lethal factors, was used in the second and third purification steps. By this means, a 105,000 mol wt. protein was purified, as judged by silver stained SDS-polyacrylamide gels. Lethal activity was inhibited by exposure to the serine protease inhibitor, L-1 chloro 3[4-tosylamido]-7-amino-2-heptanone-HCl, after purification. Although this lethal factor has some characteristics of a serine protease, it is not proteolytically active.

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Accession: 047316934

Download citation: RISBibTeXText

PMID: 7914386

DOI: 10.1016/0041-0101(94)90299-2



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