The thermal stability of the external invertase isoforms from Saccharomyces cerevisiae correlates with the surface charge density
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2012
Authors
Anđelković, Uroš
Theisgen, Stephan
Scheidt, Holger A.
Petkovic, Marijana

Huster, Daniel
Vujčić, Zoran

Article (Published version)

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Show full item recordAbstract
Understanding the effect of surface charge on the stability of proteins is one prerequisite for "tailoring" proteins with increased thermal stability. Here, we investigated the origin of the altered thermal stability observed between the four recently isolated isoforms (EINV1-EINV4) of external invertase. External invertase from yeast Saccharomyces cerevisiae, a homodimeric glycoprotein, represents a widely used model for studying the influence of the glyco component on protein stability. The stability of the four isoforms of invertase decreases from EINV1 to EINV4, which is accompanied by an increase in negative surface charge density. Mass spectrometry analysis revealed that the isoforms share identical protein parts indicating that the differences in stability are the result of post-translational modifications. P-31 NMR analysis revealed that the isoforms contain negatively charged phosphate groups in diester and monoester forms attached to the glycan part. The total amount of phosp...hate bound to the polymannan component varies between the different isoforms. These results, together with the analysis of the amount of polymannan components, show that negative surface charge density does not entirely depend on the amount of phosphate but rather on its distribution. This suggests that charged groups bound to the glyco-component of a protein can influence the stability of glycoproteins.
Keywords:
Glycoprotein stability / Surface charge / Phosphorylation / Invertase / IsoformSource:
Biochimie, 2012, 94, 2, 510-515Publisher:
- Elsevier France-Editions Scientifiques Medicales Elsevier, Paris
Funding / projects:
- Production, purification and characterization of enzymes and small molecules and their application as soluble or immobilized in food biotechnology, biofuels production and environmental protection (RS-172048)
- Deutscher Akademischer Austausch Dienst (DAAD)
DOI: 10.1016/j.biochi.2011.08.020
ISSN: 0300-9084
PubMed: 21906653
WoS: 000300270900027
Scopus: 2-s2.0-84855822854
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IHTMTY - JOUR AU - Anđelković, Uroš AU - Theisgen, Stephan AU - Scheidt, Holger A. AU - Petkovic, Marijana AU - Huster, Daniel AU - Vujčić, Zoran PY - 2012 UR - https://cer.ihtm.bg.ac.rs/handle/123456789/1014 AB - Understanding the effect of surface charge on the stability of proteins is one prerequisite for "tailoring" proteins with increased thermal stability. Here, we investigated the origin of the altered thermal stability observed between the four recently isolated isoforms (EINV1-EINV4) of external invertase. External invertase from yeast Saccharomyces cerevisiae, a homodimeric glycoprotein, represents a widely used model for studying the influence of the glyco component on protein stability. The stability of the four isoforms of invertase decreases from EINV1 to EINV4, which is accompanied by an increase in negative surface charge density. Mass spectrometry analysis revealed that the isoforms share identical protein parts indicating that the differences in stability are the result of post-translational modifications. P-31 NMR analysis revealed that the isoforms contain negatively charged phosphate groups in diester and monoester forms attached to the glycan part. The total amount of phosphate bound to the polymannan component varies between the different isoforms. These results, together with the analysis of the amount of polymannan components, show that negative surface charge density does not entirely depend on the amount of phosphate but rather on its distribution. This suggests that charged groups bound to the glyco-component of a protein can influence the stability of glycoproteins. PB - Elsevier France-Editions Scientifiques Medicales Elsevier, Paris T2 - Biochimie T1 - The thermal stability of the external invertase isoforms from Saccharomyces cerevisiae correlates with the surface charge density VL - 94 IS - 2 SP - 510 EP - 515 DO - 10.1016/j.biochi.2011.08.020 ER -
@article{ author = "Anđelković, Uroš and Theisgen, Stephan and Scheidt, Holger A. and Petkovic, Marijana and Huster, Daniel and Vujčić, Zoran", year = "2012", abstract = "Understanding the effect of surface charge on the stability of proteins is one prerequisite for "tailoring" proteins with increased thermal stability. Here, we investigated the origin of the altered thermal stability observed between the four recently isolated isoforms (EINV1-EINV4) of external invertase. External invertase from yeast Saccharomyces cerevisiae, a homodimeric glycoprotein, represents a widely used model for studying the influence of the glyco component on protein stability. The stability of the four isoforms of invertase decreases from EINV1 to EINV4, which is accompanied by an increase in negative surface charge density. Mass spectrometry analysis revealed that the isoforms share identical protein parts indicating that the differences in stability are the result of post-translational modifications. P-31 NMR analysis revealed that the isoforms contain negatively charged phosphate groups in diester and monoester forms attached to the glycan part. The total amount of phosphate bound to the polymannan component varies between the different isoforms. These results, together with the analysis of the amount of polymannan components, show that negative surface charge density does not entirely depend on the amount of phosphate but rather on its distribution. This suggests that charged groups bound to the glyco-component of a protein can influence the stability of glycoproteins.", publisher = "Elsevier France-Editions Scientifiques Medicales Elsevier, Paris", journal = "Biochimie", title = "The thermal stability of the external invertase isoforms from Saccharomyces cerevisiae correlates with the surface charge density", volume = "94", number = "2", pages = "510-515", doi = "10.1016/j.biochi.2011.08.020" }
Anđelković, U., Theisgen, S., Scheidt, H. A., Petkovic, M., Huster, D.,& Vujčić, Z.. (2012). The thermal stability of the external invertase isoforms from Saccharomyces cerevisiae correlates with the surface charge density. in Biochimie Elsevier France-Editions Scientifiques Medicales Elsevier, Paris., 94(2), 510-515. https://doi.org/10.1016/j.biochi.2011.08.020
Anđelković U, Theisgen S, Scheidt HA, Petkovic M, Huster D, Vujčić Z. The thermal stability of the external invertase isoforms from Saccharomyces cerevisiae correlates with the surface charge density. in Biochimie. 2012;94(2):510-515. doi:10.1016/j.biochi.2011.08.020 .
Anđelković, Uroš, Theisgen, Stephan, Scheidt, Holger A., Petkovic, Marijana, Huster, Daniel, Vujčić, Zoran, "The thermal stability of the external invertase isoforms from Saccharomyces cerevisiae correlates with the surface charge density" in Biochimie, 94, no. 2 (2012):510-515, https://doi.org/10.1016/j.biochi.2011.08.020 . .