Alteration of the Cell Surface during the Vegetative Cell Cycle of the Unicellular Green Alga Chlamydomonas reinhardtii

Voigt, J.; Hinkelmann, Bettina; Liebich, Ines; Mix, Marianne

doi:10.1093/oxfordjournals.pcp.a029006

Cited by 12 publications

(13 citation statements)

References 31 publications

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“…This apparent inconsistency is probably due to an oversimplification of the interpretation. Although wild-type cells are surrounded by a multilayered cell wall, this wall expands during cell growth (Voigt 1985;Voigt 1986), which transiently increases the pore size of the wall and may thereby permit transport of macromolecules, such as BSA. The cell wall barrier is composed of an insoluble, crystalline hydroxyproline-rich glycoprotein framework and several chaotrope-soluble, hydroxyproline-containing glycoproteins (Monk et al 1983).…”

Section: Discussion Electroporation Vs Sonication Mechanismsmentioning

confidence: 99%

Influence of the Cell Wall on Intracellular Delivery to Algal Cells by Electroporation and Sonication

Azencott

Peter

Prausnitz

2007

Ultrasound in Medicine & Biology

107

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To assess the cell wall's role as a barrier to intracellular delivery, wild-type Chlamydomonas reinhardtii algal cells and mutant cells lacking a cell wall were exposed to electroporation or sonication. Flow cytometry determined intracellular uptake of calcein and bovine serum albumin (BSA) and loss of cell viability as functions of electroporation transmembrane potential and acoustic energy. Electroporation of wild-type cells increased calcein uptake with increasing transmembrane potential, but delivered much less BSA. Electroporation of wall-deficient cells had similar effects on calcein uptake, but increased BSA uptake as much as 7.5-fold relative to wild-type cells, which indicated that the cell wall was a significant barrier to BSA delivery during electroporation. Sonication of wild-type cells caused calcein and BSA uptake at similar levels. This suggests that the cell wall barrier to BSA delivery can be overcome by sonication. Increased electroporation transmembrane potential or acoustic energy also caused increased loss of cell viability, where walldeficient cells were especially susceptible to lysis. Overall, we believe this is the first study to compare the effects of electroporation and sonication in a direct fashion in any cell type. Specifically, these findings suggest that electroporation primarily transports molecules across the plasma membrane, because its mechanism is specific to lipid bilayer disruption, whereas sonication transports molecules across both the plasma membrane and cell wall, because it non-specifically disrupts cell-surface barriers.

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Section: Discussion Electroporation Vs Sonication Mechanismsmentioning

confidence: 99%

Influence of the Cell Wall on Intracellular Delivery to Algal Cells by Electroporation and Sonication

Azencott

Peter

Prausnitz

2007

Ultrasound in Medicine & Biology

107

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“…The chaotrope‐soluble cell wall glycoproteins GP1, GP2, and GP3 were extracted from C. reinhardtii wild‐type cells and separated as previously described (Voigt, 1985a, b; Goodenough et al , 1986; Voigt et al , 1996, 2007).…”

Section: Methodsmentioning

confidence: 99%

The chaotrope-soluble glycoprotein GP1 is a constituent of the insoluble glycoprotein framework of theChlamydomonascell wall

Voigt¹,

Frank²,

Wöstemeyer³

2009

FEMS Microbiology Letters

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Chlamydomonas reinhardtii wild-type cells are surrounded by the insoluble cell wall component, a sac-like framework of cross-linked glycoproteins containing 22% hydroxyproline. The chaotrope-soluble cell wall glycoprotein GP1 is the only polypeptide with an even higher proportion of hydroxyproline (35%) occurring in vegetative C. reinhardtii cells. Mass spectrometric analyses of peptides released from the purified insoluble cell wall fraction by trypsin treatment and epitope analyses of polyclonal antibodies raised against different deglycosylation products of this particular wall fraction using 181 chemically synthesized GP1-derived pentadecapeptides revealed evidence that GP1 is indeed a constituent of the insoluble wall component.

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“…As reported previously, partial proteolysis of the insoluble cell wall fraction is a prerequisite for cell enlargement (Voigt, 1985(Voigt, , 1986Voigt et al, 1996) and is accompanied by a release of cell wall macromolecules into the culture medium (Voigt, 1985(Voigt, , 1986. The size of the polypeptide fragments retained in the in- Samples corresponding to 30 g of protein were fractionated by SDS-PAGE on slab gels containing 12.5% (w/v) acrylamide.…”

Section: Characterization Of Cell Wall Antibodiesmentioning

confidence: 98%

“…Therefore, enlargement of this insoluble glycoprotein framework is a prerequisite for cell growth. As revealed by pulse-labeling and pulse-chase experiments with 3 H-Pro and 35 S-Met, turnover of the insoluble framework of the Chlamydomonas cell wall takes place during the cell enlargement period accompanied by a release of large polypeptide fragments into the culture medium (Voigt, 1985(Voigt, , 1986. The resulting "holes" in the glycoprotein framework are repaired by the simultaneous incorporation of soluble precursors.…”

Section: Introductionmentioning

confidence: 99%

14-3-3 Proteins Are Constituents of the Insoluble Glycoprotein Framework of the Chlamydomonas Cell Wall

Voigt

Frank

2003

Plant Cell

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The cell wall of the unicellular green alga Chlamydomonas reinhardtii consists predominantly of Hyp-rich glycoproteins, which also occur in the extracellular matrix of multicellular green algae and higher plants. In addition to the Hyp-rich polypeptides, the insoluble glycoprotein framework of the Chlamydomonas cell wall contains minor amounts of 14-3-3 proteins, as revealed by immunochemical studies and mass spectroscopic analysis of tryptic peptides. Polypeptides immunologically related to the 14-3-3 proteins also were found in the culture medium of Chlamydomonas. The levels of two of these 14-3-3-related polypeptides were decreased in the culture medium of the wall-deficient mutant cw-15 . These findings indicate that 14-3-3 proteins are involved in the cross-linking of Hyp-rich glycoproteins in the Chlamydomonas cell wall.

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Alteration of the Cell Surface during the Vegetative Cell Cycle of the Unicellular Green Alga Chlamydomonas reinhardtii

Cited by 12 publications

References 31 publications

Influence of the Cell Wall on Intracellular Delivery to Algal Cells by Electroporation and Sonication

Influence of the Cell Wall on Intracellular Delivery to Algal Cells by Electroporation and Sonication

The chaotrope-soluble glycoprotein GP1 is a constituent of the insoluble glycoprotein framework of theChlamydomonascell wall

14-3-3 Proteins Are Constituents of the Insoluble Glycoprotein Framework of the Chlamydomonas Cell Wall

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