Document Detail

Translocation of aprotinin, a therapeutic protease inhibitor, into the thylakoid lumen of genetically engineered tobacco chloroplasts.
MedLine Citation:
PMID:  18266824     Owner:  NLM     Status:  MEDLINE    
Aprotinin, a bovine protease inhibitor of important therapeutic value, was expressed in tobacco plastid transformants. This disulphide bond-containing protein was targeted to the lumen of thylakoids using signal peptides derived from nuclear genes which encode lumenal proteins. Translocation was attempted via either the general secretion (Sec) or the twin-arginine translocation (Tat) pathway. In both cases, this strategy allowed the production of genuine aprotinin with its N-terminal arginine residue. The recombinant protease inhibitor was efficiently secreted within the lumen of thylakoids, accumulated in older leaves and was bound to trypsin, suggesting that the three disulphide bonds of aprotinin are correctly folded and paired in this chloroplast compartment. Mass spectrometric analysis indicated that translocation via the Sec pathway, unlike the Tat pathway, led predominantly to an oxidized protein. Translocation via the Tat pathway was linked to a slightly decreased growth rate, a pale-green leaf phenotype and supplementary expression products associated with the thylakoids.
Ghislaine Tissot; Hélène Canard; Marie Nadai; Aris Martone; Johan Botterman; Manuel Dubald
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Publication Detail:
Type:  Journal Article     Date:  2008-02-04
Journal Detail:
Title:  Plant biotechnology journal     Volume:  6     ISSN:  1467-7652     ISO Abbreviation:  Plant Biotechnol. J.     Publication Date:  2008 Apr 
Date Detail:
Created Date:  2008-03-07     Completed Date:  2008-05-20     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101201889     Medline TA:  Plant Biotechnol J     Country:  England    
Other Details:
Languages:  eng     Pagination:  309-20     Citation Subset:  IM    
Bayer BioScience, 69263 Lyon Cedex 09, France.
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MeSH Terms
Amino Acid Sequence
Aprotinin / genetics,  metabolism*
Chloroplasts / genetics*,  metabolism*
Gene Expression Regulation, Plant / genetics
Genetic Engineering
Plant Leaves / cytology
Plants, Genetically Modified
Protease Inhibitors / metabolism*
Protein Transport
Recombinant Proteins
Tobacco / cytology*,  genetics*
Reg. No./Substance:
0/Protease Inhibitors; 0/Recombinant Proteins; 9087-70-1/Aprotinin

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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