The Collagen V Homotrimer Production Is Unexpectedly Favored over the Heterotrimer in Recombinant Expression Systems
Figure 1
Expression of 1(V) without the native signal sequence and the C-propeptide (a, b) and of 1(V)/2(V) chains (c) in P. pastoris. (a) 5% SDS-PAGE analysis under reducing conditions of the soluble fraction of acidic extraction from untransformed yeast cells (lane 1) and from yeast cells transformed with the construct pPICZ-C5A1N (lane 2). (b) Western blot analysis of the same samples as in panel (a) probed with monoclonal pan-collagen antibody 95D1A. (c) Western blot analysis of the soluble fraction of neutral extraction of yeast cells transformed with 1(V) and 2(V) chains and induced with 0.5% methanol induction. Samples were incubated without (lane 1) or with (lane 2) pepsin before analysis. Left, molecular mass standards expressed in kDa.
