Article (Scientific journals)
Contribution of the carbohydrate moiety to conformational stability of the carboxypeptidase Y high pressure study.
Dumoulin, Mireille; Ueno, H.; Hayashi, R. et al.
1999In European Journal of Biochemistry, 262 (2), p. 475-83
Peer Reviewed verified by ORBi
 

Files


Full Text
Dumoulin_1999.pdf
Publisher postprint (715.45 kB)
Download

All documents in ORBi are protected by a user license.

Send to



Details



Keywords :
Carboxypeptidases/chemistry/metabolism; Catalysis; Cathepsin A; Enzyme Stability; Models, Molecular; Pressure; Protein Conformation; Spectrometry, Fluorescence; Temperature; Thermodynamics
Abstract :
[en] The process of pressure-induced denaturation of carboxypeptidase Y and the role of the carbohydrate moiety in its response to pressure and low temperature were investigated by measuring in situ the catalytic activity and, the intrinsic and 8-anilino-1-naphthalene sulfonic acid binding fluorescences. Pressure-induced denaturation of carboxypeptidase Y is a process involving at least three transitions. Low pressures (below 150 MPa) induced slight conformational changes characterized by a slight decrease in the center of the spectral mass of intrinsic fluorescence, whereas no changes in 8-anilino-1-naphthalene sulfonic acid binding fluorescence were observed and 80% of the catalytic activity remained. Higher pressure (150-500 MPa) induced further conformational changes, characterized by a large decrease in the center of the spectral mass of intrinsic fluorescence, a large increase in the 8-anilino-1-naphthalene sulfonic acid binding fluorescence and the loss of all catalytic activity. Thus, this intermediate exhibited characteristics of molten globule-like state. A further increase, in pressure (above 550 MPa) induced transition from this first molten globule-like state to a second molten globule-like state. This two-stage denaturation process can be explained by assuming the existence of two independent structural domains in the carboxypeptidase molecule. A similar three-transition process was found for unglycosylated carboxypeptidase Y, but, the first two transitions clearly occurred at lower pressures than those for glycosylated carboxypeptidase Y. These findings indicate that the carbohydrate moiety protects carboxypeptidase Y against pressure-induced denaturation. The origin of the protective effects is discussed based on the known crystallographic structure of CPY.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Dumoulin, Mireille  ;  Université de Liège - ULiège > Département des sciences de la vie > Enzymologie et repliement des protéines
Ueno, H.
Hayashi, R.
Balny, C.
Language :
English
Title :
Contribution of the carbohydrate moiety to conformational stability of the carboxypeptidase Y high pressure study.
Publication date :
1999
Journal title :
European Journal of Biochemistry
ISSN :
0014-2956
eISSN :
1432-1033
Publisher :
Blackwell Science, Oxford, United Kingdom
Volume :
262
Issue :
2
Pages :
475-83
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 25 November 2010

Statistics


Number of views
96 (7 by ULiège)
Number of downloads
121 (0 by ULiège)

Scopus citations®
 
36
Scopus citations®
without self-citations
21
OpenCitations
 
32

Bibliography


Similar publications



Contact ORBi