Fig. 1

Schematic representation of recombinant proteins employed in this study. All polypeptides contained human serum albumin (HSA) sequences, were expressed in Pichia pastoris, and were purified from media conditioned by methanol-induced cultures. Polypeptides are shown in linear format, amino- to carboxyl-terminus, to emphasize their modular design. Protein segments are shown in black boxes (HSA, 585 amino acids), while small proteins (HV3, hirudin variant 3, 66 amino acids, or KPI, the Kunitz Protease Inhibitor domain of protease nexin 2, 57 amino acids) are shown in grey, and peptides (G6, hexaglycine spacer, H6, hexahistidine affinity tag, EPR, FXIa-cleavable tripeptide (i.e. Glu-Pro-Arg, and myc, myc oncoprotein epitope tag, EQKLISEEDL) in white. From top to bottom, the following recombinant, His-tagged proteins are depicted, with their length in amino acids ([a.a.], at right of linear depiction): EPR-HV3HSA; HSA-EPRHV3; mycEPR-HV3HSA; mycHV3HSA; KPIHSA; and HSA