Zeolin is a recombinant storage protein with different solubility and stability properties according to its localization in the endoplasmic reticulum or in the chloroplast.
Several strategies have been exploited to maximize heterologous protein accumulation in the plant cell. Recently, it has been shown that a portion of a maize prolamin storage protein, gamma-zein, can be used for the high accumulation of a recombinant protein in novel endoplasmic reticulum (ER)-derived protein bodies of vegetative tissues. In this study, we investigate whether this protein can be expressed in the chloroplast. Our long-term purpose is to use zeolin to produce value-added proteins by fusing these polypeptides with its gamma-zein portion and targeting the recombinant proteins to the ER or to the chloroplast. We show here that zeolin accumulates in the chloroplast to lower levels than in the ER and its stability is compromised by chloroplast proteolytic activity. Co-localization of zeolin and the ER chaperone BiP in the chloroplast does not have a beneficial effect on zeolin accumulation. In this organelle, zeolin is not stored in protein bodies, nor do zeolin polypeptides seem to be linked by inter-chain disulfide bonds, which are usually formed by the six cysteine of the gamma-zein portion, indicating abnormal folding of the recombinant protein. Therefore, it is concluded that to accumulate zeolin in the chloroplast it is necessary to facilitate inter-chain disulfide bond formation.
Institute of Plant Genetics - Research Division of Perugia, Italian National Research Council (CNR), via della Madonna Alta 130, 06128 Perugia, Italy. michele.bellucci@igv.cnr.it
Michele
M
Bellucci
Francesca
F
De Marchis
Ildo
I
Nicoletti
Sergio
S
Arcioni
D002736:Chloroplasts / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified; D011489:Protein Denaturation; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D012995:Solubility; D014026:Nicotiana / Q000235:genetics; D019076:Transgenes; D015028:Zein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; C048826:phaseolin protein, Phaseolus vulgaris; D015028:Zein