AbstractA semi‐synthetic protein design approach has been employed for the structural investigation of a putative helical region at the C‐terminus of Interleukin‐2. With crystallographic or NMR derived conformational data as yet unavailable, we have relied only on primary sequence information and computer‐assisted modelling to direct the analysis. By employing both chemical peptide synthesis and recombinant DNA methods, the C‐terminus of IL‐2 was modified according to a strategy designed to stabilize helical secondary structure. A semi‐synthetic protein incorporating 12 simultaneous amino acid replacements was constructed, which possessed potentiated biological activity and displayed a far UV circular dichroism spectrum comparable to a hybrid protein with the authentic sequence. By comparison, another hybrid protein containing a C‐terminal region designed to contain helix breaking residues was totally devoid of bioactivity. These findings provide evidence that the modelling method correctly identified a helix necessary for the formation of a bioactive tertiary fold. Moreover, by employing semi‐synthesis it was possible to circumvent the difficulties associated with the preparation, purification and analysis of multiple recombinant proteins, and also to avoid the unreliability of total chemical synthesis for proteins greater