Evidence of beta-sheet structure induced kinetic stability of papain upon thermal and sodium dodecyl sulfate denaturation
Апстракт
Papain is a protease that consists of alpha-helical and beta-sheet domains that unfold almost independently. Both, considerable thermal stability and sodium dodecyl sulfate (SDS) resistance of papain have been shown. However, the ability of each domain to unfold upon thermal and SDS denaturation has never been studied. This work shows that fruit papain has slightly higher resistance to thermal inactivation when compared to that of stem papain with a rather high activation energy (E-a) of 223 +/- 16 kJ mol(-1) and a T(m)50 value of 79 +/- 2 degrees C. The SDS resistance of fruit papain was estimated by SDS-PAGE analysis and activity staining. It was noted that, in the presence of SDS the protein remained active, unless heat energy was applied in order to unfold papain. Furthermore, it was proven via Fourier transform infrared spectroscopy (FT-IR) that an alpha-helical domain of fruit papain is more prone to unfolding at elevated temperatures and in the presence of SDS then a beta-sheet ...rich domain. Thermal denaturation of papain without detergent present led to accelerated formation of aggregation specific intermolecular beta-sheets as compared to native protein. The presented results are of both fundamental and practical importance.
Кључне речи:
thermal inactivation / SDS resistance / secondary structure / FT-IR spectroscopyИзвор:
Journal of the Serbian Chemical Society, 2015, 80, 5, 613-625Издавач:
- Serbian Chemical Soc, Belgrade
Финансирање / пројекти:
- Алергени, антитела, ензими и мали физиолошки значајни молекули: дизајн, структура, функција и значај (RS-MESTD-Basic Research (BR or ON)-172049)
DOI: 10.2298/JSC140901007R
ISSN: 0352-5139
WoS: 000359987400003
Scopus: 2-s2.0-84938931203
Колекције
Институција/група
Hemijski fakultet / Faculty of ChemistryTY - JOUR AU - Rašković, Brankica AU - Babic, Nikolina AU - Korać, Jelena AU - Polović, Natalija PY - 2015 UR - https://cherry.chem.bg.ac.rs/handle/123456789/1953 AB - Papain is a protease that consists of alpha-helical and beta-sheet domains that unfold almost independently. Both, considerable thermal stability and sodium dodecyl sulfate (SDS) resistance of papain have been shown. However, the ability of each domain to unfold upon thermal and SDS denaturation has never been studied. This work shows that fruit papain has slightly higher resistance to thermal inactivation when compared to that of stem papain with a rather high activation energy (E-a) of 223 +/- 16 kJ mol(-1) and a T(m)50 value of 79 +/- 2 degrees C. The SDS resistance of fruit papain was estimated by SDS-PAGE analysis and activity staining. It was noted that, in the presence of SDS the protein remained active, unless heat energy was applied in order to unfold papain. Furthermore, it was proven via Fourier transform infrared spectroscopy (FT-IR) that an alpha-helical domain of fruit papain is more prone to unfolding at elevated temperatures and in the presence of SDS then a beta-sheet rich domain. Thermal denaturation of papain without detergent present led to accelerated formation of aggregation specific intermolecular beta-sheets as compared to native protein. The presented results are of both fundamental and practical importance. PB - Serbian Chemical Soc, Belgrade T2 - Journal of the Serbian Chemical Society T1 - Evidence of beta-sheet structure induced kinetic stability of papain upon thermal and sodium dodecyl sulfate denaturation VL - 80 IS - 5 SP - 613 EP - 625 DO - 10.2298/JSC140901007R ER -
@article{ author = "Rašković, Brankica and Babic, Nikolina and Korać, Jelena and Polović, Natalija", year = "2015", abstract = "Papain is a protease that consists of alpha-helical and beta-sheet domains that unfold almost independently. Both, considerable thermal stability and sodium dodecyl sulfate (SDS) resistance of papain have been shown. However, the ability of each domain to unfold upon thermal and SDS denaturation has never been studied. This work shows that fruit papain has slightly higher resistance to thermal inactivation when compared to that of stem papain with a rather high activation energy (E-a) of 223 +/- 16 kJ mol(-1) and a T(m)50 value of 79 +/- 2 degrees C. The SDS resistance of fruit papain was estimated by SDS-PAGE analysis and activity staining. It was noted that, in the presence of SDS the protein remained active, unless heat energy was applied in order to unfold papain. Furthermore, it was proven via Fourier transform infrared spectroscopy (FT-IR) that an alpha-helical domain of fruit papain is more prone to unfolding at elevated temperatures and in the presence of SDS then a beta-sheet rich domain. Thermal denaturation of papain without detergent present led to accelerated formation of aggregation specific intermolecular beta-sheets as compared to native protein. The presented results are of both fundamental and practical importance.", publisher = "Serbian Chemical Soc, Belgrade", journal = "Journal of the Serbian Chemical Society", title = "Evidence of beta-sheet structure induced kinetic stability of papain upon thermal and sodium dodecyl sulfate denaturation", volume = "80", number = "5", pages = "613-625", doi = "10.2298/JSC140901007R" }
Rašković, B., Babic, N., Korać, J.,& Polović, N.. (2015). Evidence of beta-sheet structure induced kinetic stability of papain upon thermal and sodium dodecyl sulfate denaturation. in Journal of the Serbian Chemical Society Serbian Chemical Soc, Belgrade., 80(5), 613-625. https://doi.org/10.2298/JSC140901007R
Rašković B, Babic N, Korać J, Polović N. Evidence of beta-sheet structure induced kinetic stability of papain upon thermal and sodium dodecyl sulfate denaturation. in Journal of the Serbian Chemical Society. 2015;80(5):613-625. doi:10.2298/JSC140901007R .
Rašković, Brankica, Babic, Nikolina, Korać, Jelena, Polović, Natalija, "Evidence of beta-sheet structure induced kinetic stability of papain upon thermal and sodium dodecyl sulfate denaturation" in Journal of the Serbian Chemical Society, 80, no. 5 (2015):613-625, https://doi.org/10.2298/JSC140901007R . .