Producing recombinant Crotalus adamanteus Snake venom metalloproteinase adamalysin-2 first isolates the target gene, which covers the full-length Snake venom metalloproteinase adamalysin-2 (1-203aa). This gene is co-cloned into an appropriate expression vector with an N-terminal 6xHis-tag gene and introduced into E. coli cells. The positive E. coli cells are cultured to express the recombinant protein. After that, these cells undergo lysis to release the protein. Purification of the protein is typically achieved using affinity chromatography. The purity of the recombinant Snake venom metalloproteinase adamalysin-2 is over 90% as determined by SDS-PAGE.
Crotalus adamanteus adamalysin-2 is a significant component found in the venom of the Crotalus adamanteus. This protein belongs to the adamalysin or ADAM family, which includes proteolytic domains found in snake venom metalloproteinases, mammalian reproductive tract proteins, and tumor necrosis factor-alpha convertase (TACE) [1]. Adamalysin-2 is a 24 kDa zinc endopeptidase that is part of a large family of metalloproteinases and is crucial for various biological activities, requiring Zn2+ and Ca2+ as cofactors [2]. It is a well-known metalloendopeptidase toxin that can be inhibited by certain molecules like Jatromollistatin, a cyclic heptapeptide, as demonstrated through molecular docking analysis [3].
References:
[1] F. Gomis‐Rüth, E. Meyer, L. Kress, & V. Politi, Structures of adamalysin ii with peptidic inhibitors. implications for the design of tumor necrosis factor α convertase inhibitors, Protein Science, vol. 7, no. 2, p. 283-292, 1998. https://doi.org/10.1002/pro.5560070207
[2] P. Phan, A review of rattlesnake venoms, Toxins, vol. 16, no. 1, p. 2, 2023. https://doi.org/10.3390/toxins16010002
[3] T. Jucá, M. Ramos, E. Cilli, A. Neto, S. Mackessy, & A. Monteiro-Moreira, Insights on the inhibition properties ofjatromollistatin(a cyclic heptapeptide) againstcrotalus adamanteusmetalloendopeptidase using molecular docking analysis, Journal of Molecular Recognition, vol. 35, no. 7, 2022. https://doi.org/10.1002/jmr.2957
