Use of endogenous signal sequences for transient production and efficient secretion by moss (Physcomitrella patens) cells.
BACKGROUND
Efficient targeting to appropriate cell organelles is one of the bottlenecks for the production of recombinant proteins in plant systems. A common practice is to use the native secretory signal peptide of the heterologous protein to be produced. Though general features of secretion signals are conserved between plants and animals, the broad sequence variability among signal peptides suggests differing efficiency of signal peptide recognition.


RESULTS
Aiming to improve secretion in moss bioreactors, we quantitatively compared the efficiency of two human signal peptides and six signals from recently isolated moss (Physcomitrella patens) proteins. We therefore used fusions of the different signals to heterologous reporter sequences for transient transfection of moss cells and measured the extra- and intracellular accumulation of the recombinant proteins rhVEGF and GST, respectively. Our data demonstrates an up to fivefold higher secretion efficiency with endogenous moss signals compared to the two utilised human signal peptides.


CONCLUSIONS
From the distribution of extra- and intracellular recombinant proteins, we suggest translational inhibition during the signal recognition particle-cycle (SRP-cycle) as the most probable of several possible explanations for the decreased extracellular accumulation with the human signals. In this work, we report on the supremacy of moss secretion signals over the utilised heterologous ones within the moss-bioreactor system. Though the molecular details of this effect remain to be elucidated, our results will contribute to the improvement of molecular farming systems.
Schaaf
Andreas
A
Department of Plant Biochemistry and Biotechnology, University of Münster, Hindenburgplatz 55, 48143 Münster, Germany. schaaf@uni-muenster.de
Tintelnot
Stefanie
S
Baur
Armin
A
Reski
Ralf
R
Gorr
Gilbert
G
Decker
Eva L
EL
D000595:Amino Acid Sequence; D019149:Bioreactors; D001709:Biotechnology / Q000379:methods*; D044002:Bryophyta; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D017930:Genes, Reporter; D005821:Genetic Techniques; D005822:Genetic Vectors; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D021382:Protein Sorting Signals; D011994:Recombinant Proteins / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D042461:Vascular Endothelial Growth Factor A; D005982:Glutathione Transferase