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

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

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

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
Pages (from-to)131-141
Number of pages11
JournalPlant Science
Volume162
Issue number1
DOIs
StatePublished - 2002

Fingerprint

gamma-zein
Zein
disulfide bonds
Disulfides
Zea mays
Cysteine
cysteine
corn
Prolamins
Solubility
prolamins
Proteins
solubility
proteins
Fusion reactions
protein folding
protein solubility
Protein folding
Thioredoxins
sulfides

Keywords

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

ASJC Scopus subject areas

  • Plant Science
  • Biochemistry
  • Biotechnology

Cite this

Mutational analysis of the maize gamma zein C-terminal cysteine residues. / Ems-McClung, Stephanie C.; Benmoussa, Mustapha; Hainline, Bryan.

In: Plant Science, Vol. 162, No. 1, 2002, p. 131-141.

Research output: Contribution to journalArticle

Ems-McClung, Stephanie C. ; Benmoussa, Mustapha ; Hainline, Bryan. / Mutational analysis of the maize gamma zein C-terminal cysteine residues. In: Plant Science. 2002 ; Vol. 162, No. 1. pp. 131-141.
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