Determination of the disulfide bond pairings in bovine transforming growth factor-alpha |
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| Authors: | B N Violand J S Tou B D Vineyard N R Siegel C E Smith P D Pyla J F Zobel P C Toren E W Kolodziej |
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| Affiliation: | Animal Sciences Division, Monsanto Company, St. Louis, MO. |
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| Abstract: | A rapid method for determining the three disulfide bond pairings in bovine transforming growth factor-alpha (bTGF-alpha) was developed by digesting bTGF-alpha 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-alpha were determined to be homologous to those in the human and mouse TGF-alpha molecules. A species of low bioactivity isolated from the folding/oxidation mixture of chemically synthesized bTGF-alpha was demonstrated to contain two incorrect disulfide bonds. These results indicate that mispairing of disulfide bonds in bTGF-alpha significantly reduces the activity of this molecule. |
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