Molecular mass determination and assay of venom hyaluronidases by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
We describe a procedure for molecular mass determination of hyaluronidases present in animal venoms from different families. Hyaluronidases can be revealed, following their electrophoretic separation in sodium dodecyl sulfate-polyacrylamide gel containing hyaluronic acid, by incubating the gel in Triton X-100 to remove sodium dodecyl sulfate and restore in situ enzyme activity. This method allows the detection of as little as 0.025 turbidity-reducing units after 2 hr incubation. All the hyaluronidases from the analyzed invertebrate venoms had a mass below 50,000 and showed only one component, while those from vertebrate venoms were more than 60,000 and in many instances contained more than one form.
Cevallos
M A
MA
Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Mor.
Navarro-Duque
C
C
Varela-Julia
M
M
Alagon
A C
AC
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006821:Hyaluronoglucosaminidase / Q000032:analysis*; D008970:Molecular Weight; D012967:Sodium Dodecyl Sulfate; D014688:Venoms / Q000737:chemistry* / Q000201:enzymology / Q000302:isolation & purification
D014688:Venoms; D012967:Sodium Dodecyl Sulfate; D006821:Hyaluronoglucosaminidase