Home LiteratureArticle Details
PMID: 16632592 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The intracellular fate of a recombinant protein is tissue dependent.

Plant physiology ·Vol. 141 ·No. 2 ·2006-06-00 ·Pages 578-86

Drakakaki G, Marcel S, Arcalis E, Altmann F, Gonzalez-Melendi P, Fischer R, Christou P, Stoger E

Abstract

Recombinant proteins directed to the secretory pathway in plants require a signal peptide for entry into the endoplasmic reticulum. In the absence of further targeting information, such proteins are generally secreted via the default pathway to the apoplast. This has been well documented in protoplasts and leaf tissue, but the trafficking of recombinant proteins in seeds and other storage tissues has rarely been investigated. We used Aspergillus niger phytase as a model glycoprotein to compare the intracellular fate of a recombinant protein in the leaves and seeds of rice (Oryza sativa). Using fluorescence and electron microscopy we showed that the recombinant protein was efficiently secreted from leaf cells as expected. In contrast, within endosperm cells it was retained in endoplasmic reticulum-derived prolamin bodies and protein storage vacuoles. Consistent with our immunolocalization data, the phytase produced in endosperm cells possessed oligomannose and vacuolar-type N-glycans [Man(3)(Xyl)(Fuc)GlcNAc(2)], whereas the phytase produced in leaves contained predominantly secretion-type N-glycans [GlcNAc(2)Man(3)(Xyl)(Fuc)GlcNAc(2)]. The latter could not be detected in preparations of the endosperm-derived phytase. Our results show that the intracellular deposition and modification of a recombinant protein is tissue dependent.

MeSH Terms
6-Phytase/chemistry,genetics Base Sequence DNA Primers Endoplasmic Reticulum/metabolism In Situ Hybridization Microscopy, Electron Microscopy, Fluorescence Molecular Weight Oryza/enzymology,genetics Plant Leaves/metabolism Plant Proteins/metabolism Plants, Genetically Modified RNA, Messenger/genetics Recombinant Proteins/metabolism Seeds/metabolism
Chemicals
DNA Primers Plant Proteins RNA, Messenger Recombinant Proteins 6-Phytase
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Drakakaki Georgia
Institute for Molecular Biotechnology, Biology VII, Aachen University, 52074 Aachen, Germany.
Marcel Sylvain
Arcalis Elsa
Altmann Friedrich
Gonzalez-Melendi Pablo
Fischer Rainer
Christou Paul
Stoger Eva
References (46)
46 references, click to expand
  1. Transport of storage proteins to protein storage vacuoles is mediated by large precursor-accumulating vesicles
    Plant Cell. 1998 May;10(5):825-36 PMID: 9596640
  2. Cereal seed storage proteins: structures, properties and role in grain utilization.
    J Exp Bot. 2002 Apr;53(370):947-58 PMID: 11912237
  3. Differential targeting of the tobacco PR-1 pathogenesis-related proteins to the extracellular space and vacuoles of crystal idioblasts.
    EMBO J. 1991 Jun;10(6):1317-24 PMID: 2026137
  4. Extracellular secretion of Aspergillus phytase from Arabidopsis roots enables plants to obtain phosphorus from phytate.
    Plant J. 2001 Mar;25(6):641-9 PMID: 11319031
  5. Deposition of storage proteins.
    Plant Mol Biol. 1998 Sep;38(1-2):77-99 PMID: 9738961
  6. Sorting of proteins to vacuoles in plant cells.
    Plant Mol Biol. 1998 Sep;38(1-2):127-44 PMID: 9738964
  7. N-glycoprotein biosynthesis in plants: recent developments and future trends.
    Plant Mol Biol. 1998 Sep;38(1-2):31-48 PMID: 9738959
  8. N-Glycan analysis by matrix-assisted laser desorption/ionization mass spectrometry of electrophoretically separated nonmammalian proteins: application to peanut allergen Ara h 1 and olive pollen allergen Ole e 1.
    Anal Biochem. 2000 Oct 1;285(1):64-75 PMID: 10998264
  9. Protein glycosylation. Structural and functional aspects.
    Eur J Biochem. 1993 Nov 15;218(1):1-27 PMID: 8243456
  10. Endoplasmic reticulum to vacuole trafficking of endoplasmic reticulum bodies provides an alternate pathway for protein transfer to the vacuole.
    Plant Physiol. 2004 Nov;136(3):3440-6 PMID: 15542498
  11. Cloned and expressed fungal phyA gene in alfalfa produces a stable phytase.
    Biochem Biophys Res Commun. 2002 Feb 1;290(4):1343-8 PMID: 11812011
  12. Expression and localization of human lysozyme in the endosperm of transgenic rice.
    Planta. 2003 Feb;216(4):597-603 PMID: 12569401
  13. A recombinant multimeric immunoglobulin expressed in rice shows assembly-dependent subcellular localization in endosperm cells.
    Plant Biotechnol J. 2005 Jan;3(1):115-27 PMID: 17168904
  14. Mass transport of proform of a KDEL-tailed cysteine proteinase (SH-EP) to protein storage vacuoles by endoplasmic reticulum-derived vesicle is involved in protein mobilization in germinating seeds.
    J Cell Biol. 2000 Feb 7;148(3):453-64 PMID: 10662772
  15. Unexpected deposition patterns of recombinant proteins in post-endoplasmic reticulum compartments of wheat endosperm.
    Plant Physiol. 2004 Nov;136(3):3457-66 PMID: 15489278
  16. Improvement of in-gel digestion protocol for peptide mass fingerprinting by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.
    Rapid Commun Mass Spectrom. 2001;15(16):1416-21 PMID: 11507753
  17. Rice prolamine protein body biogenesis: a BiP-mediated process.
    Science. 1993 Nov 12;262(5136):1054-6 PMID: 8235623
  18. Localization of 5 S genes and transcripts in Pisum sativum nuclei.
    J Cell Sci. 1993 Aug;105 ( Pt 4):1151-8 PMID: 8227204
  19. Protein secretion in plant cells can occur via a default pathway.
    Plant Cell. 1990 Jan;2(1):51-9 PMID: 1967050
  20. Sorting of proteins to storage vacuoles: how many mechanisms?
    Trends Plant Sci. 2005 Jul;10(7):316-23 PMID: 15950520
  21. Development of biopharmaceuticals in plant expression systems: cloning, expression and immunological reactivity of human cytomegalovirus glycoprotein B (UL55) in seeds of transgenic tobacco.
    Vaccine. 1999 Aug 6;17(23-24):3020-9 PMID: 10462237
  22. Stable accumulation of Aspergillus niger phytase in transgenic tobacco leaves.
    Plant Physiol. 1995 Dec;109(4):1199-205 PMID: 8539288
  23. Messenger RNA targeting of rice seed storage proteins to specific ER subdomains.
    Nature. 2000 Oct 12;407(6805):765-7 PMID: 11048726
  24. Rice cell culture as an alternative production system for functional diagnostic and therapeutic antibodies.
    Transgenic Res. 1999;8(6):441-9 PMID: 10767987
  25. A novel vesicle derived directly from endoplasmic reticulum is involved in the transport of vacuolar storage proteins in rice endosperm.
    Plant Cell Physiol. 2005 Jan;46(1):245-9 PMID: 15659439
  26. Fungal phyA gene expressed in potato leaves produces active and stable phytase.
    Biochem Biophys Res Commun. 2003 Jun 27;306(2):603-9 PMID: 12804608
  27. Dual regulated RNA transport pathways to the cortical region in developing rice endosperm.
    Plant Cell. 2003 Oct;15(10):2265-72 PMID: 14523246
  28. Cloning, characterization and overexpression of the phytase-encoding gene (phyA) of Aspergillus niger.
    Gene. 1993 May 15;127(1):87-94 PMID: 8387447
  29. COMPARTMENTATION OF PROTEINS IN THE ENDOMEMBRANE SYSTEM OF PLANT CELLS.
    Annu Rev Plant Physiol Plant Mol Biol. 1996 Jun;47:327-350 PMID: 15012292
  30. Evidence for a novel route of wheat storage proteins to vacuoles.
    J Cell Biol. 1992 Dec;119(5):1117-28 PMID: 1447291
  31. Immunochemical studies on the role of the Golgi complex in protein-body formation in rice seeds.
    Planta. 1986 Dec;169(4):471-80 PMID: 24232753
  32. Protein sorting to the storage vacuoles of plants: a critical appraisal.
    Traffic. 2005 Aug;6(8):615-25 PMID: 15998318
  33. Where, oh where has my protein gone?
    Trends Biotechnol. 2004 Feb;22(2):53-5 PMID: 14757034
  34. The transport of prolamine RNAs to prolamine protein bodies in living rice endosperm cells.
    Plant Cell. 2003 Oct;15(10):2253-64 PMID: 14508010
  35. N-glycosylation potential of maize: the human lactoferrin used as a model.
    Glycoconj J. 2001 Jul;18(7):519-27 PMID: 12151713
  36. [Expression of phytase gene in Bombyx mori].
    Sheng Wu Gong Cheng Xue Bao. 2003 Jan;19(1):112-5 PMID: 15969047
  37. Analysis of Asn-linked glycans from vegetable foodstuffs: widespread occurrence of Lewis a, core alpha1,3-linked fucose and xylose substitutions.
    Glycobiology. 2001 Apr;11(4):261-74 PMID: 11358875
  38. A long and winding road: symposium on membrane trafficking in plants.
    EMBO Rep. 2004 Mar;5(3):245-9 PMID: 14993924
  39. Localization of a bacterial protein in starch granules of transgenic maize kernels.
    Proc Natl Acad Sci U S A. 2003 Sep 16;100(19):11127-32 PMID: 12960364
  40. Comparative analysis of the site-specific N-glycosylation of human lactoferrin produced in maize and tobacco plants.
    Eur J Biochem. 2003 Aug;270(15):3235-42 PMID: 12869199
  41. Targeting and release of phytohemagglutinin from the roots of bean seedlings.
    Plant Physiol. 1995 Oct;109(2):603-10 PMID: 7480348
  42. The cargo in vacuolar storage protein transport vesicles is stratified.
    Traffic. 2005 Jan;6(1):45-55 PMID: 15569244
  43. Sorting of glycoprotein B from human cytomegalovirus to protein storage vesicles in seeds of transgenic tobacco.
    Transgenic Res. 2001 Apr;10(2):177-81 PMID: 11305363
  44. Secretion of active recombinant phytase from soybean cell-suspension cultures.
    Plant Physiol. 1997 Jul;114(3):1103-11 PMID: 9232886
  45. Targeting of proteins to endoplasmic reticulum-derived compartments in plants. The importance of RNA localization.
    Plant Physiol. 2004 Nov;136(3):3414-9 PMID: 15542494
  46. Expression of an Aspergillus niger phytase gene (phyA) in Saccharomyces cerevisiae.
    Appl Environ Microbiol. 1999 May;65(5):1915-8 PMID: 10223979
Article Info
Journal
Plant physiology
Abbr.
Plant Physiol
ISSN
0032-0889
DOI
10.1104/pp.106.076661
Published
2006-06-00
Epub
2006-00-21
Pages
578-86
Language
English
Region
United States
NLM ID
0401224
PMCID
PMC1475444
Subset
IM
External Links
PubMed source DOI: 10.1104/pp.106.076661 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2