Infolding and refolding of active apple polyphenol oxidase
For the first time, unfolding (6 M guanidine) and refolding of partially proteolysed purified polyphenol oxidase (PPOr) was achieved, with 88% of activity recovered. Optimal refolding conditions consisted in stepwise dialysis of guanidine treated extracts, the dialysis buffers containing 1 M (NH4)2SO4 and 100 [mu]M CuSO4. However, CuSO4 had limited effect on the recovering of PPOr activity, whereas (NH4)2SO4 was essential. Concerning the PPO tertiary structure, denaturing conditions (combinations of boiling and reducing agent) used on SDS-PAGE have shown (i) a compact tertiary structure and (ii) the presence of disulfide bonds in PPOr, accounting for the shift between 27 and 41 kDa, and 41 and 42 kDa, respectively. Resistance to proteolytic cleavage was used to study the conformational changes induced by the denaturing treatments. Folded PPOr was resistant to further proteolysis whereas unfolded PPO was totally digested, indicating the role of tertiary structure of PPOr in the resistance to proteases.
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dig-cirad-fr-5252722024-06-27T16:01:36Z http://agritrop.cirad.fr/525272/ http://agritrop.cirad.fr/525272/ Infolding and refolding of active apple polyphenol oxidase. Mari Stéphane, Marquès Laurence, Breton Frédéric, Karamanos Yannis, Macheix Jean-Jacques. 1998. Phytochemistry, 49 (5) : 1213-1217.https://doi.org/10.1016/S0031-9422(98)00142-3 <https://doi.org/10.1016/S0031-9422(98)00142-3> Infolding and refolding of active apple polyphenol oxidase Mari, Stéphane Marquès, Laurence Breton, Frédéric Karamanos, Yannis Macheix, Jean-Jacques eng 1998 Phytochemistry For the first time, unfolding (6 M guanidine) and refolding of partially proteolysed purified polyphenol oxidase (PPOr) was achieved, with 88% of activity recovered. Optimal refolding conditions consisted in stepwise dialysis of guanidine treated extracts, the dialysis buffers containing 1 M (NH4)2SO4 and 100 [mu]M CuSO4. However, CuSO4 had limited effect on the recovering of PPOr activity, whereas (NH4)2SO4 was essential. Concerning the PPO tertiary structure, denaturing conditions (combinations of boiling and reducing agent) used on SDS-PAGE have shown (i) a compact tertiary structure and (ii) the presence of disulfide bonds in PPOr, accounting for the shift between 27 and 41 kDa, and 41 and 42 kDa, respectively. Resistance to proteolytic cleavage was used to study the conformational changes induced by the denaturing treatments. Folded PPOr was resistant to further proteolysis whereas unfolded PPO was totally digested, indicating the role of tertiary structure of PPOr in the resistance to proteases. article info:eu-repo/semantics/article Journal Article info:eu-repo/semantics/publishedVersion http://agritrop.cirad.fr/525272/1/525272.pdf text Cirad license info:eu-repo/semantics/restrictedAccess https://agritrop.cirad.fr/mention_legale.html https://doi.org/10.1016/S0031-9422(98)00142-3 10.1016/S0031-9422(98)00142-3 http://catalogue-bibliotheques.cirad.fr/cgi-bin/koha/opac-detail.pl?biblionumber=185423 info:eu-repo/semantics/altIdentifier/doi/10.1016/S0031-9422(98)00142-3 info:eu-repo/semantics/altIdentifier/purl/https://doi.org/10.1016/S0031-9422(98)00142-3 |
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For the first time, unfolding (6 M guanidine) and refolding of partially proteolysed purified polyphenol oxidase (PPOr) was achieved, with 88% of activity recovered. Optimal refolding conditions consisted in stepwise dialysis of guanidine treated extracts, the dialysis buffers containing 1 M (NH4)2SO4 and 100 [mu]M CuSO4. However, CuSO4 had limited effect on the recovering of PPOr activity, whereas (NH4)2SO4 was essential. Concerning the PPO tertiary structure, denaturing conditions (combinations of boiling and reducing agent) used on SDS-PAGE have shown (i) a compact tertiary structure and (ii) the presence of disulfide bonds in PPOr, accounting for the shift between 27 and 41 kDa, and 41 and 42 kDa, respectively. Resistance to proteolytic cleavage was used to study the conformational changes induced by the denaturing treatments. Folded PPOr was resistant to further proteolysis whereas unfolded PPO was totally digested, indicating the role of tertiary structure of PPOr in the resistance to proteases. |
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article |
author |
Mari, Stéphane Marquès, Laurence Breton, Frédéric Karamanos, Yannis Macheix, Jean-Jacques |
spellingShingle |
Mari, Stéphane Marquès, Laurence Breton, Frédéric Karamanos, Yannis Macheix, Jean-Jacques Infolding and refolding of active apple polyphenol oxidase |
author_facet |
Mari, Stéphane Marquès, Laurence Breton, Frédéric Karamanos, Yannis Macheix, Jean-Jacques |
author_sort |
Mari, Stéphane |
title |
Infolding and refolding of active apple polyphenol oxidase |
title_short |
Infolding and refolding of active apple polyphenol oxidase |
title_full |
Infolding and refolding of active apple polyphenol oxidase |
title_fullStr |
Infolding and refolding of active apple polyphenol oxidase |
title_full_unstemmed |
Infolding and refolding of active apple polyphenol oxidase |
title_sort |
infolding and refolding of active apple polyphenol oxidase |
url |
http://agritrop.cirad.fr/525272/ http://agritrop.cirad.fr/525272/1/525272.pdf |
work_keys_str_mv |
AT maristephane infoldingandrefoldingofactiveapplepolyphenoloxidase AT marqueslaurence infoldingandrefoldingofactiveapplepolyphenoloxidase AT bretonfrederic infoldingandrefoldingofactiveapplepolyphenoloxidase AT karamanosyannis infoldingandrefoldingofactiveapplepolyphenoloxidase AT macheixjeanjacques infoldingandrefoldingofactiveapplepolyphenoloxidase |
_version_ |
1806036906855628800 |