Substrate zymography of snake venom components depicting the activity of matrix metalloprotease

Abstract

Introduction: Matrix metalloproteases (MMPs) are a group of endopeptidases belonging to a superfamily of zinc protease enzymes that have the potential to degrade the extracellular matrix (ECM). The loss of its activity could lead to very serious alterations in the human body, causing several conditions such as arthritis and aneurisms. Previous studies indicate that snake venom contains snake venom metalloprotease (SVMP), which might possess similar activity to MMPs. In these experiments, it was shown that Malaysian cobra SVMP is capable of gelatinolytic activity, which was demonstrated using gelatine as substrate in sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) (zymography). Objectives: To investigate the activity of Malaysian cobra SVMP using zymography. Materials and methods: Protein was extracted from cobra snake venom using fast protein liquid chromatography then, after purification, the fractions were collected and run on SDS-PAGE to visualize the protein of interest, which was then desalted and concentrated using dialysis and ready to be used for zymography to demonstrate the activity of SVMP. Results: In the detection of the SVMP on SDS-PAGE, the molecular weight was observed around 55kDa. A possible dimer was also seen at 110kDa. Gelatinase activity of SVMP was clearly revealed on the gelatine gel that was used in zymography. Conclusions: Having found the gelatinolytic activity of SVMP the future direction of this protein can be tested against the tumour environment to see the potential use of SVMP’ s. Acknowledgements: Al-Imam Muhammad ibn Saud Islamic University for their support, International Medical University for using their laboratories, and Dr Fabian Davamani for supervising this project.