Determination of Disulfide Bonds in gamma-46 Gliadin

T. A. Egorov^1*, T. I. Odintsova², and A. K. Musolyamov¹

¹Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, ul. Vavilova 32, Moscow, 117984 Russia; fax: (095) 135-1405; E-mail: egorov@imb.imb.ac.ru

²Vavilov Institute of General Genetics, Russian Academy of Sciences, ul. Gubkina 3, Moscow, 117809 Russia

^* To whom correspondence should be addressed.

Received July 3, 1998; Revision received December 3, 1998

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The disulfide bonds in gamma-46 gliadin were identified: Cys₁₇₃--Cys₁₉₂, Cys₂₁₂--Cys₂₉₁, Cys₁₆₅--Cys₁₉₉ (or Cys₂₀₀), Cys₂₈₃--Cys₂₀₀ (or Cys₁₉₉). The disulfide-containing peptides were obtained by limited hydrolysis of the intact protein with chymotrypsin at an enzyme/substrate ratio of 1:1000 at 20°C for 22 h with subsequent digestion of disulfide-containing fragments with trypsin and chymotrypsin. The locations of disulfide bonds were determined by sequencing disulfide-containing fractions and constituent peptides and comparison of the obtained sequences with the partial amino acid sequence of gamma-46 gliadin determined earlier.

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KEY WORDS: storage proteins, prolamins, gliadins, disulfide bonds

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