Recombinant collagen 8A1(Col8A1) is expressed and purified from HEK293 cells. It runs at ~250kDa under non-reducing condition on SDS-PAGE, but why is it dissociating into a single chain (70kDa) on a DTT reduced gel since there are no disulfide bonds?
Dear Wei Li ,
It depends on what you mean with non-reducing (no DTT obviously but also no boiling and presence of urea?).
Indeed, it has nothing to do with disulphide bonds, but everything with the ability of collagen to form (homo)trimers. See for more details: https://www.uniprot.org/uniprot/P27658
Best regards.
PS. In the mentioned link they refer to the following paper:
Stephan, S., Sherratt, M. J., Hodson, N., Shuttleworth, C. A., & Kielty, C. M. (2004). Expression and supramolecular assembly of recombinant α1 (VIII) and α2 (VIII) collagen homotrimers. Journal of Biological Chemistry, 279(20), 21469-21477.
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