Determination of the disulfide bond pairings in bovine transforming growth factor‐α

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Abstract
A rapid method for determining the three disulfide bond pairings in bovine transforming growth factor‐α (bTGF‐α) was developed by digesting bTGF‐α with thermolysin followed by separation of the generated peptides by reversed‐phase HPLC. The disulfide‐bonded peptides were identified by amino acid sequencing and fast atom bombardment mass spectrometry. The disulfide bond pairings in bTGF‐α were determined to be homologous to those in the human and mouse TGF‐α molecules. A species of low bioactivity isolated from the folding/oxidation mixture of chemically synthesized bTGF‐α was demonstrated to contain two incorrect disulfide bonds. These results indicate that mispairing of disulfide bonds in bTGF‐α significantly reduces the activity of this molecule.