Effect of acetonitrile on Cynara cardunculus L. cardosin A stability

Anna V. Shnyrova; Claudia S. Oliveira; Ana C. Sarmento; Marlene T. Barros; Galina G. Zhadan; Manuel G. Roig; Valery L. Shnyrov

doi:10.1016/j.ijbiomac.2006.04.007

Effect of acetonitrile on Cynara cardunculus L. cardosin A stability

Anna V. Shnyrova, Claudia S. Oliveira, Ana C. Sarmento, Marlene T. Barros, Galina G. Zhadan, Manuel G. Roig, Valery L. Shnyrov^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

5 Citations (Scopus)

Abstract

The kinetics of the structural changes affecting cardosin A, a plant aspartic proteinase (AP) from Cynara cardunculus L., in the presence of a mixture of acetonitrile (AN) in water (W) was studied. Incubation of cardosin A with 10% (v/v) AN resulted in a gradual increase in protein helicity, accompanied by changes in the tertiary structure, seen by changes in the intrinsic fluorescence of tryptophan. Differential scanning calorimetry (DSC) revealed that the temperature of denaturation of cardosin A decreased upon the addition of AN. With longer incubation times, the small chain of cardosin A denatured completely, consequent exposure of the single tryptophan residue accounting well for the observed spectral shift intrinsic fluorescence of the protein. Enzymatic activity assays demonstrated that the kinetically determined unfolding of the small chain of cardosin A does not result in loss of the activity of this enzyme.

Original language	English
Pages (from-to)	273-279
Number of pages	7
Journal	International Journal of Biological Macromolecules
Volume	39
Issue number	4-5
DOIs	https://doi.org/10.1016/j.ijbiomac.2006.04.007
Publication status	Published - 15 Nov 2006

Keywords

Cardosin A
Differential scanning calorimetry
Low organic medium catalysis
Protein stability
Water-acetonitrile mixture

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10.1016/j.ijbiomac.2006.04.007

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title = "Effect of acetonitrile on Cynara cardunculus L. cardosin A stability",

abstract = "The kinetics of the structural changes affecting cardosin A, a plant aspartic proteinase (AP) from Cynara cardunculus L., in the presence of a mixture of acetonitrile (AN) in water (W) was studied. Incubation of cardosin A with 10% (v/v) AN resulted in a gradual increase in protein helicity, accompanied by changes in the tertiary structure, seen by changes in the intrinsic fluorescence of tryptophan. Differential scanning calorimetry (DSC) revealed that the temperature of denaturation of cardosin A decreased upon the addition of AN. With longer incubation times, the small chain of cardosin A denatured completely, consequent exposure of the single tryptophan residue accounting well for the observed spectral shift intrinsic fluorescence of the protein. Enzymatic activity assays demonstrated that the kinetically determined unfolding of the small chain of cardosin A does not result in loss of the activity of this enzyme.",

keywords = "Cardosin A, Differential scanning calorimetry, Low organic medium catalysis, Protein stability, Water-acetonitrile mixture",

author = "Shnyrova, {Anna V.} and Oliveira, {Claudia S.} and Sarmento, {Ana C.} and Barros, {Marlene T.} and Zhadan, {Galina G.} and Roig, {Manuel G.} and Shnyrov, {Valery L.}",

year = "2006",

month = nov,

day = "15",

doi = "10.1016/j.ijbiomac.2006.04.007",

language = "English",

volume = "39",

pages = "273--279",

journal = "International Journal of Biological Macromolecules",

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T1 - Effect of acetonitrile on Cynara cardunculus L. cardosin A stability

AU - Shnyrova, Anna V.

AU - Oliveira, Claudia S.

AU - Sarmento, Ana C.

AU - Barros, Marlene T.

AU - Zhadan, Galina G.

AU - Roig, Manuel G.

AU - Shnyrov, Valery L.

PY - 2006/11/15

Y1 - 2006/11/15

N2 - The kinetics of the structural changes affecting cardosin A, a plant aspartic proteinase (AP) from Cynara cardunculus L., in the presence of a mixture of acetonitrile (AN) in water (W) was studied. Incubation of cardosin A with 10% (v/v) AN resulted in a gradual increase in protein helicity, accompanied by changes in the tertiary structure, seen by changes in the intrinsic fluorescence of tryptophan. Differential scanning calorimetry (DSC) revealed that the temperature of denaturation of cardosin A decreased upon the addition of AN. With longer incubation times, the small chain of cardosin A denatured completely, consequent exposure of the single tryptophan residue accounting well for the observed spectral shift intrinsic fluorescence of the protein. Enzymatic activity assays demonstrated that the kinetically determined unfolding of the small chain of cardosin A does not result in loss of the activity of this enzyme.

AB - The kinetics of the structural changes affecting cardosin A, a plant aspartic proteinase (AP) from Cynara cardunculus L., in the presence of a mixture of acetonitrile (AN) in water (W) was studied. Incubation of cardosin A with 10% (v/v) AN resulted in a gradual increase in protein helicity, accompanied by changes in the tertiary structure, seen by changes in the intrinsic fluorescence of tryptophan. Differential scanning calorimetry (DSC) revealed that the temperature of denaturation of cardosin A decreased upon the addition of AN. With longer incubation times, the small chain of cardosin A denatured completely, consequent exposure of the single tryptophan residue accounting well for the observed spectral shift intrinsic fluorescence of the protein. Enzymatic activity assays demonstrated that the kinetically determined unfolding of the small chain of cardosin A does not result in loss of the activity of this enzyme.

KW - Cardosin A

KW - Differential scanning calorimetry

KW - Low organic medium catalysis

KW - Protein stability

KW - Water-acetonitrile mixture

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DO - 10.1016/j.ijbiomac.2006.04.007

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AN - SCOPUS:33750181319

SN - 0141-8130

VL - 39

SP - 273

EP - 279

JO - International Journal of Biological Macromolecules

JF - International Journal of Biological Macromolecules

IS - 4-5

ER -

Effect of acetonitrile on Cynara cardunculus L. cardosin A stability

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Keywords

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