Mutational analysis of the maize gamma zein C-terminal cysteine residues

Stephanie C. Ems-McClung, Mustapha Benmoussa, Bryan E. Hainline

Research output: Contribution to journalArticle

6 Scopus citations


Alignment of 98 S-rich prolamin amino acid sequences illustrated that four cysteines were preserved nearly 100% and four cysteines were less well conserved. These residues form two to four intramolecular disulfide bonds in several S-rich prolamins. For maize gamma zein, it is unknown whether the eight C-terminal cysteines form intramolecular disulfide bonds. If gamma zein contains intramolecular disulfide bonds, modifications of critical cysteines in soluble thioredoxin-gamma zein fusion protein might prevent disulfide bond formation and result in misfolded and insoluble protein in Escherichia coli. Individual modification of conserved cysteines C128 and C136 resulted in a four-fold reduction in solubility and a significant decrease in expression level compared to wild-type fusion protein. Modification of conserved C156 resulted in a two-fold reduction in expression level but not a significant change in solubility until combined with a non-deleterious Q181R modification. This suggested that C128 and C136 were involved in disulfide bonds critical for protein folding, whereas C156 was more critical for protein stability. Modification of a non-conserved N-terminal cysteine residue (C117) resulted in increased protein solubility, suggesting it was not involved in an intramolecular disulfide bond. From these data and a review of the literature, a disulfide map for gamma zein is proposed.

Original languageEnglish (US)
Pages (from-to)131-141
Number of pages11
JournalPlant Science
Issue number1
StatePublished - Jan 8 2002


  • Cysteine
  • Gamma zein
  • Genetic engineering
  • Intramolecular disulfide bonds
  • Phenylketonuria

ASJC Scopus subject areas

  • Plant Science
  • Biochemistry
  • Biotechnology

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