Comparative properties of amplified external and internal invertase from the yeast SUC2 gene.

Williams, Richard S.; Trumbly, Robert; MacColl, Robert; Trimble, Robert B.; Maley, Frank

doi:10.1016/s0021-9258(17)38874-9

Cited by 45 publications

(5 citation statements)

References 34 publications

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“…The role of carbohydrate associated with external invertase has been the subject of several investigations, which in essence have revealed that the oligosaccharides are not essential for enzyme activity (Trimble & Maley, 1977;Tarentino et al, 1974) or for the maintenance of the peptide backbone conformation (Williams et al, 1985). In addition, denaturation studies have shown that the oligosaccharides do not afford protection against GuHCl or heat (Trumbly et al, 1985;Shulte & Schmid, 1988a,b).…”

Section: Discussionmentioning

confidence: 99%

“…External invertase was purchased from Boehringer and Mannheim (specific activity about 300 units/mg of protein) and separated from internal invertase and contaminating carbohydrate material by column chromatography. Further purification on a Sephacryl S-300 (Pharmacia) column yielded the final preparation (specific activity 4500 units/mg of protein) which was homogeneous on SDS-PAGE (Trimble & Maley, 1977;Williams et al, 1985).…”

Section: Methodsmentioning

confidence: 99%

“…Cloned internal invertase was purified from yeast cells by ion-exchange, hydrophobic, and gel filtration chromatographies on DE-52, phenyl-Sepharose CL-4B (Pharmacia), and Ultrogel AcA44 (LKB) columns, respectively (Williams et al, 1985). The final preparation had a specific activity comparable to external invertase and was homogeneous on SDS-PAGE.…”

Section: Methodsmentioning

confidence: 99%

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Effect of oligosaccharides and chloride on the oligomeric structures of external, internal, and deglycosylated invertase

Reddy¹,

MacColl²,

Maley³

1990

Biochemistry

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Effect of oligosaccharides and chloride on the oligomeric structures of external, internal, and deglycosylated invertase

Reddy¹,

MacColl²,

Maley³

1990

Biochemistry

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“…To test the specificity of signal peptide-independent translocation, we examined Suc2DN20, an N-terminal signal peptidedeprived derivative of Suc2 (extracellular invertase) and Png1 (cytosolic protein) containing no intrinsic signal peptide (25,26). Both proteins have potential N-glycosylation sites.…”

Section: Resultsmentioning

confidence: 99%

The ER-associated protease Ste24 prevents N-terminal signal peptide-independent translocation into the endoplasmic reticulum in Saccharomyces cerevisiae

Hosomi

Iida

Cho

et al. 2020

Journal of Biological Chemistry

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Soluble proteins destined for the secretory pathway contain an N-terminal signal peptide that induces their translocation into the endoplasmic reticulum (ER). The importance of N-terminal signal peptides for ER translocation has been extensively examined over the past few decades. However, in the budding yeast Saccharomyces cerevisiae, a few proteins devoid of a signal peptide are still translocated into the ER and then N-glycosylated. Using signal peptide–truncated reporter proteins, we herein report detection of significant translocation of N-terminal signal peptide–truncated proteins in a yeast mutant strain (ste24∆) that lacks the endopeptidase Ste24 at the ER membrane. Furthermore, several ER/cytosolic proteins, including Sec61, Sec66, and Sec72, were identified as being involved in the translocation process. On the basis of screening for 20 soluble proteins that may be N-glycosylated in the ER in the ste24∆ strain, we identified the transcription factor Rme1 as a protein that is partially N-glycosylated despite the lack of a signal peptide. These results clearly indicate that some proteins lacking a signal peptide can be translocated into the ER and that Ste24 typically suppresses this process.

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“…However, yeast S. cerevisiae is the main source of invertase in the food industry. S. cerevisiae synthesizes two forms of invertase with the same amino acid sequence: the external (glycosylated) and the internal (nonglycosylated) , . The external invertase is of industrial importance due to the presence of a carbohydrate component, which increases its thermal stability , resistance against protease , and solubility , and makes the enzyme extremely stable at room temperature .…”

Section: Introductionmentioning

confidence: 99%

Thermodynamics and Structural Features of the YeastSaccharomyces cerevisiaeExternal Invertase Isoforms in Guanidinium-chloride Solutions

Andjelković

Lah

2010

J. Agric. Food Chem.

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Recently, four external invertase isoforms (EINV1, EINV2, EINV3, and EINV4) have been isolated from S. cerevisiae. However, there is nothing known about their structural features and thermodynamics of unfolding. Since this information is essential for understanding their functioning at the molecular level as well as applicable in the food industry, we investigated guanidinium-chloride induced structural changes of the isoforms by CD and fluorescence spectroscopy. The resulting unfolding curves measured for each isoform at different temperatures were described simultaneously by a reversible two-state model to obtain the corresponding thermodynamic parameters. Here, we show that they are different for different isoforms and demonstrate that they correlate with the surface charge density of the native isoforms which follows the order EINV1 < EINV2 < EINV3 < EINV4. It appears that at physiological temperatures the thermodynamic stability of the isoforms follows the same order, while above 55 °C, the order is the opposite EINV1 > EINV2 > EINV3 ≈ EINV4. This suggests that increasing the efficiency of the food industry processes involving invertase would require the application of EINV3 and/or EINV4 at physiological temperatures and EINV1 at elevated temperatures.

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Comparative properties of amplified external and internal invertase from the yeast SUC2 gene.

Cited by 45 publications

References 34 publications

Effect of oligosaccharides and chloride on the oligomeric structures of external, internal, and deglycosylated invertase

Effect of oligosaccharides and chloride on the oligomeric structures of external, internal, and deglycosylated invertase

The ER-associated protease Ste24 prevents N-terminal signal peptide-independent translocation into the endoplasmic reticulum in Saccharomyces cerevisiae

Thermodynamics and Structural Features of the YeastSaccharomyces cerevisiaeExternal Invertase Isoforms in Guanidinium-chloride Solutions

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