Refolding of a carboxypeptidase Y folding intermediate in vitro by low-affinity binding of the proregion

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Refolding of a carboxypeptidase Y folding intermediate in vitro by low-affinity binding of the proregion. / Winther, Jakob R.; Sørensen, P; Kielland-Brandt, Morten.

In: Journal of Biological Chemistry, Vol. 269, No. 35, 1994, p. 22007-13.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Winther, JR, Sørensen, P & Kielland-Brandt, M 1994, 'Refolding of a carboxypeptidase Y folding intermediate in vitro by low-affinity binding of the proregion', Journal of Biological Chemistry, vol. 269, no. 35, pp. 22007-13.

APA

Winther, J. R., Sørensen, P., & Kielland-Brandt, M. (1994). Refolding of a carboxypeptidase Y folding intermediate in vitro by low-affinity binding of the proregion. Journal of Biological Chemistry, 269(35), 22007-13.

Vancouver

Winther JR, Sørensen P, Kielland-Brandt M. Refolding of a carboxypeptidase Y folding intermediate in vitro by low-affinity binding of the proregion. Journal of Biological Chemistry. 1994;269(35):22007-13.

Author

Winther, Jakob R. ; Sørensen, P ; Kielland-Brandt, Morten. / Refolding of a carboxypeptidase Y folding intermediate in vitro by low-affinity binding of the proregion. In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 35. pp. 22007-13.

Bibtex

@article{aabc268c932f4e0eadcc984507534b31,
title = "Refolding of a carboxypeptidase Y folding intermediate in vitro by low-affinity binding of the proregion",
abstract = "Efficient folding of carboxypeptidase Y is dependent on the presence of the proregion. Thus, denatured procarboxypeptidase Y, in contrast to the mature enzyme, refolds efficiently in vitro in low ionic strength buffers. Under these conditions denatured mature carboxypeptidase Y forms an inactive, soluble folding intermediate, which has been characterized in the present study. The inactive intermediate can be folded into the active enzyme at a low efficiency (5-10%) by the addition of 0.9 M ammonium sulfate. The refolding is accompanied by pronounced structural changes. As seen for other protease zymogens the isolated proregion from carboxypeptidase Y was found to stimulate refolding without covalent linkage to the mature part. However, the added proregion does not form a stable complex with the native enzyme and requires the presence of 0.9 M ammonium sulfate to exhibit its function. The proregion increases the yield of correctly folded enzyme, and kinetic analysis suggests that this is due to a reduction of the rate of nonproductive folding or aggregation. In addition, the proregion stabilizes carboxypeptidase Y toward thermoinactivation.",
keywords = "Carboxypeptidases, Cathepsin A, Chromatography, Gel, Electrophoresis, Polyacrylamide Gel, Kinetics, Protein Binding, Protein Folding, Protein Precursors, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins",
author = "Winther, {Jakob R.} and P S{\o}rensen and Morten Kielland-Brandt",
year = "1994",
language = "English",
volume = "269",
pages = "22007--13",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology, Inc.",
number = "35",

}

RIS

TY - JOUR

T1 - Refolding of a carboxypeptidase Y folding intermediate in vitro by low-affinity binding of the proregion

AU - Winther, Jakob R.

AU - Sørensen, P

AU - Kielland-Brandt, Morten

PY - 1994

Y1 - 1994

N2 - Efficient folding of carboxypeptidase Y is dependent on the presence of the proregion. Thus, denatured procarboxypeptidase Y, in contrast to the mature enzyme, refolds efficiently in vitro in low ionic strength buffers. Under these conditions denatured mature carboxypeptidase Y forms an inactive, soluble folding intermediate, which has been characterized in the present study. The inactive intermediate can be folded into the active enzyme at a low efficiency (5-10%) by the addition of 0.9 M ammonium sulfate. The refolding is accompanied by pronounced structural changes. As seen for other protease zymogens the isolated proregion from carboxypeptidase Y was found to stimulate refolding without covalent linkage to the mature part. However, the added proregion does not form a stable complex with the native enzyme and requires the presence of 0.9 M ammonium sulfate to exhibit its function. The proregion increases the yield of correctly folded enzyme, and kinetic analysis suggests that this is due to a reduction of the rate of nonproductive folding or aggregation. In addition, the proregion stabilizes carboxypeptidase Y toward thermoinactivation.

AB - Efficient folding of carboxypeptidase Y is dependent on the presence of the proregion. Thus, denatured procarboxypeptidase Y, in contrast to the mature enzyme, refolds efficiently in vitro in low ionic strength buffers. Under these conditions denatured mature carboxypeptidase Y forms an inactive, soluble folding intermediate, which has been characterized in the present study. The inactive intermediate can be folded into the active enzyme at a low efficiency (5-10%) by the addition of 0.9 M ammonium sulfate. The refolding is accompanied by pronounced structural changes. As seen for other protease zymogens the isolated proregion from carboxypeptidase Y was found to stimulate refolding without covalent linkage to the mature part. However, the added proregion does not form a stable complex with the native enzyme and requires the presence of 0.9 M ammonium sulfate to exhibit its function. The proregion increases the yield of correctly folded enzyme, and kinetic analysis suggests that this is due to a reduction of the rate of nonproductive folding or aggregation. In addition, the proregion stabilizes carboxypeptidase Y toward thermoinactivation.

KW - Carboxypeptidases

KW - Cathepsin A

KW - Chromatography, Gel

KW - Electrophoresis, Polyacrylamide Gel

KW - Kinetics

KW - Protein Binding

KW - Protein Folding

KW - Protein Precursors

KW - Saccharomyces cerevisiae

KW - Saccharomyces cerevisiae Proteins

M3 - Journal article

C2 - 8071321

VL - 269

SP - 22007

EP - 22013

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 35

ER -

ID: 43974389