TY - JOUR
T1 - Comparative study of mature and zymogen mite cysteine protease stability and pH unfolding
AU - Chevigné, Andy
AU - Dumez, Marie Eve
AU - Dumoulin, Mireille
AU - Matagne, André
AU - Jacquet, Alain
AU - Galleni, Moreno
N1 - Funding Information:
This work was supported by the Belgian Fonds de la Recherche Scientifique (FRS-FNRS) and the Fonds de la Recherche and Fondamentale et Collective (2.4.524.03, 2.4.511.06), IUAP P6/19-. AC is Aspirant of the FRS (Brussels, Belgium) and ME D is supported by the FRIA (Brussels, Belgium). AM is Research associate of the FRS-FNRS and is supported in part by a grant from the “Fonds de la Recherche Fondamentale et Collective” (contract number 2.4550.05). MD is Research associate of the FRS-FNRS.
PY - 2010/9
Y1 - 2010/9
N2 - Background: Papain-like proteases (CA1) are synthesized as inactive precursors carrying an N-terminal propeptide, which is further removed under acidic conditions to generate active enzymes. Methods: To have a better insight into the mechanism of activation of this protease family, we compared the pH unfolding of the zymogen and the mature form of the mite cysteine protease Der p 1. Results: We showed that the presence of the propeptide does not significantly influence the pH-induced unfolding of the catalytic domain but does affect its fluorescence properties by modifying the exposure of the tryptophan 192 to the solvent. In addition, we demonstrated that the propeptide displays weaker pH stability than the protease domain confirming that the unfolding of the propeptide is the key event in the activation process of the zymogen. General significance: Finally, we show, using thermal denaturation and enzymatic activity measurements, that whatever the pH value, the propeptide does not stabilize the structure of the catalytic domain but very interestingly, prevents its autolysis.
AB - Background: Papain-like proteases (CA1) are synthesized as inactive precursors carrying an N-terminal propeptide, which is further removed under acidic conditions to generate active enzymes. Methods: To have a better insight into the mechanism of activation of this protease family, we compared the pH unfolding of the zymogen and the mature form of the mite cysteine protease Der p 1. Results: We showed that the presence of the propeptide does not significantly influence the pH-induced unfolding of the catalytic domain but does affect its fluorescence properties by modifying the exposure of the tryptophan 192 to the solvent. In addition, we demonstrated that the propeptide displays weaker pH stability than the protease domain confirming that the unfolding of the propeptide is the key event in the activation process of the zymogen. General significance: Finally, we show, using thermal denaturation and enzymatic activity measurements, that whatever the pH value, the propeptide does not stabilize the structure of the catalytic domain but very interestingly, prevents its autolysis.
KW - Allergen
KW - Cysteine protease
KW - Der p 1
KW - Fluorescence quenching
KW - Mechanism of activation
KW - PH-induced unfolding
KW - Propeptide
KW - Stability
KW - Thermal denaturation
KW - Zymogen
UR - http://www.scopus.com/inward/record.url?scp=77955282029&partnerID=8YFLogxK
U2 - 10.1016/j.bbagen.2010.05.011
DO - 10.1016/j.bbagen.2010.05.011
M3 - Article
C2 - 20682463
AN - SCOPUS:77955282029
SN - 0304-4165
VL - 1800
SP - 937
EP - 945
JO - Biochimica et Biophysica Acta - General Subjects
JF - Biochimica et Biophysica Acta - General Subjects
IS - 9
ER -