BioMed Research International / 2010 / Article / Fig 1

Research Article

The Collagen V Homotrimer Production Is Unexpectedly Favored over the Heterotrimer in Recombinant Expression Systems

Figure 1

Expression of 𝛼 1(V) without the native signal sequence and the C-propeptide (a, b) and of 𝛼 1(V)/ 𝛼 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 𝛼 -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 𝛼 1(V) and 𝛼 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.

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