Comparison of cell wall proteins of Saccharomyces cerevisiae as anchors for cell surface expression of heterologous proteins

Vaart, J.M. van der; Biesebeke, Rob te; Chapman, John W.; Toschka, Holger Y.; Klis, Frans M.; Verrips, C. Theo

doi:10.1128/aem.63.2.615-620.1997

Cited by 135 publications

(53 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Bacteria replicate autonomously, in contrast to bacteriophages, and are sufficiently large to be analyzed by optical methods, including fluorescence microscopy, or high-throughput methods, such as fluorescence-activated cell sorting (FACS) (13,20,21,24,46,52,142). Since the first reports of bacterial display of heterologous proteins in 1986 by Freudl and coworkers (26) and Charbit and coworkers (17), an extraordinary number of different display systems has been established for yeasts (15,106,129,132,149), grampositive bacteria (40,69,84,102,117,119,123), and gramnegative bacteria (1,25,27,53,56,59,63,71,72,112,118,141). These systems have been employed for a wide range of biotechnical and biomedical applications and have prompted substantial progress in whole-cell biocatalysis, live vaccine development, biosorbent and biosensor development, epitope mapping, antigen delivery, inhibitor design, and protein/peptide library screening (for overviews, see references 5, 28, 67, and 143).…”

Section: Introductionmentioning

confidence: 99%

The Autodisplay Story, from Discovery to Biotechnical and Biomedical Applications

Jose

Meyer

2007

Microbiol Mol Biol Rev

192

189

View full text Add to dashboard Cite

SUMMARY Among the pathways used by gram-negative bacteria for protein secretion, the autotransporter pathway represents a solution of impressive simplicity. Proteins are transported, independent of their nature as recombinant or native passengers, as long as the coding nucleotide sequence is inserted in frame between those of an N-terminal signal peptide and a C-terminal domain, referred to as the β-barrel of the outer membrane translocation unit. The immunoglobulin A1 (IgA1) protease from Neisseria gonorrhoeae was the first identified member of the autotransporter family of secreted proteins. The IgA1 protease was employed in initial experiments investigating autotransporter-mediated surface display of recombinant proteins and to investigate structural and functional requirements. Various other autotransporter proteins have since been described, and the autodisplay system was developed on the basis of the natural Escherichia coli autotransporter protein AIDA-I (adhesin involved in diffuse adherence). Autodisplay has been used for the surface display of random peptide libraries to successfully screen for novel enzyme inhibitors. The autodisplay system was also used for the surface display of functional enzymes, including esterases, oxidoreductases, and electron transfer proteins. Whole E. coli cells displaying enzymes have been utilized to efficiently synthesize industrially important rare organic compounds with specific chirality. Autodisplay of epitopes on the surface of attenuated Salmonella carriers has also provided a novel way to induce immune protection after oral vaccination. This review summarizes the structural and functional features of the autodisplay system, illustrating its discovery and most recent applications. Autodisplay facilitates the export of more than 100,000 recombinant molecules per single cell and permits the oligomerization of subunits on the cell surface as well as the incorporation of inorganic prosthetic groups after transport of apoproteins onto the bacterial surface without disturbing bacterial integrity or viability. We discuss future biotechnical and biomedical applications in the light of these achievements.

show abstract

Section: Introductionmentioning

confidence: 99%

The Autodisplay Story, from Discovery to Biotechnical and Biomedical Applications

Jose

Meyer

2007

Microbiol Mol Biol Rev

192

189

View full text Add to dashboard Cite

show abstract

“…However, the dibasic motif generally found in the GPI-anchored plasma membrane protein of S. cerevisiae was not found in HpGas1p, indicating a slight difference in the GPIanchoring motifs between two strains. Such a difference helps to explain the fact that S. cerevisiae Cwp2p, which was the most effective anchoring motif in S. cerevisiae (van der Vaart et al, 1997), was less effective in H. polymorpha compared to other endogenous anchors (Table 2).…”

Section: Discussionmentioning

confidence: 99%

“…Vectors containing the 40 amino acids of the anchoring motif were named Cwp40, Tip40, Gas40, and Sed40. As a control surface anchoring motif, a PCR fragment containing the 68 C-terminal amino acids of the S. cerevisiae Cwp2p protein (van der Vaart et al, 1997) was also subcloned into the BamHI/HindIII site of pGK-CMCF and named ScCwp2. All plasmids were introduced into the H. polymorpha DL1-L (leu2 ) and LEU + transformants were selected on minimal selective plate.…”

Section: Evaluation Of Gpi-anchored Proteins As a Surface Display Motifmentioning

confidence: 99%

“…The Tip1p of S. cerevisiae (ScTip1p) is a GPIanchored cell wall protein consisting of 210 amino acids, encoded by the TIP1 gene, showing a high degree of homology with Tir1p (GenBank Accession No. M71216; van der Vaart et al, 1995). A putative HpTIP1 ORF with 283 amino acids (GenBank Accession No.…”

Section: Cloning Of Genes Encoding the Gpi-anchored Proteinmentioning

confidence: 99%

“…Cwp1p is a typical GPI-anchored cell wall protein that is encoded by the 720 bp CWP1 gene of S. cerevisiae (GenBank Accession No. D37975;van der Vaart et al, 1995;Shimoi et al, 1995). The cloned Sal I fragment contained several ORFs, one of which comprised 81 amino acids with a potential cell wall protein structure.…”

Section: Cloning Of Genes Encoding the Gpi-anchored Proteinmentioning

confidence: 99%

See 2 more Smart Citations

A cell surface display system using novel GPI‐anchored proteins in Hansenula polymorpha

Kim

Sohn

Pyun

et al. 2002

Yeast

View full text Add to dashboard Cite

A cell surface display system was developed in yeast Hansenula polymorpha.The four genes HpSED1, HpGAS1, HpTIP1 and HpCWP1, encoding glycosylphosphatidylinositol (GPI)-anchored cell surface proteins from H. polymorpha, were cloned, characterized and evaluated for their efficacies as cell surface display motifs of reporter proteins. Sequence analysis of these genes revealed that each encodes a typical GPIanchored protein that is structurally similar to a counterpart gene in S. cerevisiae. The genes showed a high content of serine-threonine (alanine) and harboured a putative secretion signal in the N-terminus and the GPI-attachment signal in the C-terminus. The surface anchoring efficiency of these putative cell surface proteins was tested by fusion to the C-terminal of carboxymethylcellulase (CMCase) from Bacillus subtilis. In all cases, high CMCase activities were detected in intact cell fraction, indicating anchoring of CMCase to the cell surface. HpCwp1p, HpGas1p and the 40 C-terminal amino acids of HpTip1p from H. polymorphaexhibited a comparatively high CMCase surface anchoring efficiency. When these proteins were used as anchoring motifs for surface display of the glucose oxidase (GOD) from Aspergillus niger, most enzyme activity was detected at the cell surface. Fluorescence activated cell sorter (FACS) analysis of cells displaying GOD on the cell surface demonstrated that GOD was well exposed on the cell surface. HpCwp1p showed the highest anchoring efficiency among others.

show abstract

In silicio identification of glycosyl-phosphatidylinositol-anchored plasma-membrane and cell wall proteins ofSaccharomyces cerevisiae

Caro

Tettelin

Vossen

et al. 1997

Yeast

314

325

View full text Add to dashboard Cite

Use of the Von Heijne algorithm allowed the identification of 686 open reading frames (ORFs) in the genome of Saccharomyces cerevisiae that encode proteins with a potential N-terminal signal sequence for entering the secretory pathway. On further analysis, 51 of these proteins contain a potential glycosyl-phosphatidylinositol (GPI)attachment signal. Seven additional ORFs were found to belong to this group. Upon examination of the possible GPI-attachment sites, it was found that in yeast the most probable amino acids for GPI-attachment are asparagine and glycine.In yeast, GPI-proteins are found at the cell surface, either attached to the plasma-membrane or as an intrinsic part of the cell wall. It was noted that plasma-membrane GPI-proteins possess a dibasic residue motif just before their predicted GPI-attachment site. Based on this, and on homologies between proteins, families of plasma-membrane and cell wall proteins were assigned, revealing 20 potential plasma-membrane and 38 potential cell wall proteins. For members of three plasma-membrane protein families, a function has been described. On the other hand, most of the cell wall proteins seem to be structural components of the wall, responsive to different growth conditions.The GPI-attachment site of yeast slightly differs from mammalian cells. This might be of use in the development of anti-fungal drugs. 1997 John Wiley & Sons, Ltd.

show abstract

Comparison of cell wall proteins of Saccharomyces cerevisiae as anchors for cell surface expression of heterologous proteins

Cited by 135 publications

References 33 publications

The Autodisplay Story, from Discovery to Biotechnical and Biomedical Applications

The Autodisplay Story, from Discovery to Biotechnical and Biomedical Applications

A cell surface display system using novel GPI‐anchored proteins in Hansenula polymorpha

In silicio identification of glycosyl-phosphatidylinositol-anchored plasma-membrane and cell wall proteins ofSaccharomyces cerevisiae

Contact Info

Product

Resources

About